Background Investigating the association of blue light-induced damage of retinal pigmenepithelial (RPE) cellwith intracellulaCa2+ conteniimportanfounderstanding the mechanism of retinal disorders.Objective Thistudy wato establish blue light-induced damage model of human RPE celland explore the relationship between the damage of RPE cell and intracellulaCa2+ content.MethodHuman RPE cellwere cultured and passaged.Cell vitality waassayed by trypan blue staining.Fourth-generation cellwere used in these experiments.The cellwere exposed to blue lighwith an intensity of (2000±500)lx fo3,6,9 o12 hours,and the rate of apoptosiwaassayed by TUNEL to assesthe optimal irradiation time focellcultured.The cellwere then randomized into the withouirradiation group,irradiation only group,nifedipine group,ligh+ nifedipine group,(-) BayK8644 group and ligh+ (-) BayK8644 group.The laseconfocal microscope waused to determine the fluorescence intensity of intracellulafree Ca2+ aan excitation wavelength of 488 nm and an emission wavelength of 505 nm.The cell imagewere analyzed using computesoftware.The differenceof fluorescence intensity among the differengroupwere compared by one-way analysiof variance.ResultTrypan blue staining showed thathe viability of RPE cellwamore than 90％ afteculturing and passaging.No apoptoticell waseen aftelighexposure fo3 hours.However,differennumberof apoptoticellappeared aftelighexposure fo6,9 and 12 hours.The fluorescence intensity of intracellulafree Ca2+ in the nifedipine group wasignificantly lowethan thaof the withouirradiation group othe ligh+ nifedipine group(both aP=0.000).Lasescanning confocal microscopy showed thathe fluorescence intensitieof intracellulafree Ca2+ in the irradiation only group,(-) BayK8644 group,ligh+ (-)BayK8644 group were highethan thaof the withouirradiation group,with statistical significancebetween them(all P=0.000).No significandifferencewere found in the fluorescence intensity of intracellulafree Ca2+ between the ligh+ nifedipine group and withouirradiation group awell abetween the (-)BayK8644 group and the ligh+(-) BayK8644 group(P =0.339,P =0.410).ConclusionThe optical conditionfoblue light-induced RPE cell damage were exposure of blue-ligha(2000± 500) lx fo6 hourand culturing the cellfo24 hours.Blue lighexposure can induce damage of human RPE cellprobably by triggering the increase of intracellulafree Ca2+ concentration.

OBJECTIVETo establish an appropriate animal model of uterine leiomyoma and to understand the pathogenesis of this disease.

METHODSMature female rats were intramuscularly injected with estradiol benzoate at 200 μg or 300 μg twice a week. After injection for 8 or 10 weeks, the rats were sacrificed. We measured the serum levels of estrogen (E(2)) and progesterone (P), evaluated ER and PR expression, and calculated the leiomyoma forming rate and mortality of the rats. Histological changes were compared between rat uterine leiomyoma and human uterine leiomyoma with HE staining. The optimal dose and duration of E(2) for induction of uterine leiomyoma in rat were determined.

RESULTSIn the rats treated with estradiol benzoate 200 μg for 8 weeks ìn the serum E(2) level increased significantly (P<0.01). Uterine nodules were visible in some of the tested rats. Based on the pathohistological Results , the uterine leiomyoma developed in the treated rats demonstrated similar features as in human uterine leiomyoma. The expressions of ER and PR were increased in the leiomyoma tissues.

CONCLUSIONThe rat model of uterine leiomyoma can be established by intramuscular injection of estradiol benzoate at 200 μg twice per week for 8 weeks, with similar features as those of human uterine leiomyoma. The high concentrations of ER and PR in uterine tissue might be related with the development of uterine leiomyoma in animal.

Animals ; Disease Models, Animal ; Estrogens ; administration & dosage ; adverse effects ; Female ; Leiomyoma ; chemically induced ; Rats ; Uterine Neoplasms ; chemically induced

OBJECTIVETo analyze clinical features, therapeutic effects and prognostic factors of patients with mantle cell lymphoma (MCL).

METHODSClinical data of 37 MCL patients hospitalized in our hospital from January 2000 to March 2010 were retrospectively analyzed.

RESULTSThe median age was 62, with a male predominance. 97.30% of the patients were in Ann Arbor stage III ∼ IV, 54.05% with B symptoms, 64.86% with bone marrow involvement, 29.73% with splenomegaly, 24.32% with lymphocytosis and 51.35% with elevated LDH. Ki-67 was detected in 22 cases, and patients with Ki-67 ≤ 40% accounted for 68.18%. Of 37 cases, the overall response rate (ORR) of rituximab combined with chemotherapy was 92.31%, being higher than those of CHOP (46.15%) and CHOP + IFN (42.86%) regimens. There were statistical differences in the 3-year progression-free survival (PFS) and overall survival (OS) between rituximab + chemotherapy and CHOP or CHOP + interferon regimens (P < 0.05, respectively). Splenomegaly, elevated WBC, lymphocytosis and Ki-67 > 40% were identified as adverse prognostic factors.

CONCLUSIONMost patients with MCL were older adults, with a male predominance and usually had bone marrow involvement and poor prognosis. Rituximab combined with chemotherapy could improve ORR and OS of MCL.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Disease-Free Survival ; Female ; Humans ; Lymphoma, Mantle-Cell ; diagnosis ; drug therapy ; mortality ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Survival Analysis

OBJECTIVETo study the severity of periodontitis and risk factors in Chengdu.

METHODS202 periodontitis patients (65 male, 137 female), aged from 25 to 60, were requested to fill a questionnaire. Probing depth (PD), clinical attachment level (CAL), gingival recession and bleeding on probing (BOP) on 6 sites of each tooth were measured and recorded.

RESULTSThe mean PD, AL, gingival recession and BOP% of 202 subjects was (3.2 +/- 0.31) mm, (3.5 +/- 0.37) mm, (0.3 +/- 0.02) mm and 21.16%. 59% of subjects missed at least one tooth. 129 subjects suffered with initial to moderate periodontitis. 73 subject suffered with advanced periodontitis. 40, 86, 55 and 21 subjects had received college education, high school education, middle school education and primary school education. 18% of subjects had smoking history, 67% subjects had tea/coffee history, 66% of subjects had psychosocial problem, and only 8% of subjects had received regular periodontal treatment. There is no relationship between the severity of periodontitis and education.

CONCLUSIONIt is very important to develop an education program on oral healthy for people in Chengdu.

Adult ; Female ; Gingival Recession ; Humans ; Male ; Middle Aged ; Periodontal Attachment Loss ; Periodontal Index ; Periodontitis ; Risk Factors

OBJECTIVETo explore the role of glial cell metabolism in the generation and regulation of central respiratory rhythm.

METHODSThe medulla oblongata slices (600-700 microm) containing the medial region of the nucleus retrofacialis (mNRF) with the hypoglossal nerve rootlets retained from 12 neonatal (0-3 days) Sprague-Dawley rats were prepared and perfused with modified Kreb's solution (MKS). Upon recording of respiratory rhythmical discharge activity (RRDA) of the rootlets of the hypoglossal nerve, the brain slices were treated with glial cell metabolism antagonist L-methionine sulfoximine (L-MSO, 50 micromol/L) for 20 min followed by application of glial cell metabolism agonist L-glutamine (L-GLN, 30 micromol/L) for 20 min, or with L-MSO for 20 min with additional L-GLN for 20 min. The changes in the RRDA of the rootlets of the hypoglossal nerve in response to the treatments were recorded.

RESULTSL-MSO prolonged the respiratory cycle (RC) and expiratory time (TE), and reduced the integral amplitude (IA) and the inspiratory time (TI) in the brain slices. L-GLN induced a significant decrease in RC and TE, but IA and TI showed no obvious variations. The effect of L-MSO on the respiratory rhythm was reversed by the application of L-GLN.

CONCLUSIONGlial cell metabolism may play an important role in the modulation of RRDA in neonatal rat brainstem.

Animals ; Animals, Newborn ; Glutamine ; pharmacology ; In Vitro Techniques ; Medulla Oblongata ; metabolism ; physiology ; Methionine Sulfoximine ; pharmacology ; Neuroglia ; metabolism ; Periodicity ; Rats ; Rats, Sprague-Dawley ; Respiration

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 explore the role of group II metabotropic glutamate receptors in the modulation of basic respiratory rhythm.

METHODSNeonatal (0-3 days) SD rats of either sex were used. The medulla oblongata brain slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared, and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O2 and 5% CO2) within 3 min. The brain slices were quickly transferred to a recording chamber and continuously perfused with oxygen-saturated MKS at a rate of 4-6 ml/min at 27-29 degrees celsius. Eighteen medulla oblongata slices were divided into 3 groups and treated for 10 min with group II metabotropic glutamate receptor-specific agonist 2R,4R-4-aminopyrrolidine-2,4-dicarboxylate (APDC) (at concentrations of 10, 20, 50 micromol/L), group II metabotropic glutamate receptor antagonist (2S)-alpha-ethylglutamic acid (EGLU) (300 micromol/L), or APDC (50 micromol/L)+EGLU (300 micromol/L) after a 10 min APDC (50 micromol/L) application. Respiratory rhythmical discharge activity (RRDA) of the rootlets of the hypoglossal nerve was recorded by suction electrodes.

RESULTSAPDC produced a dose-dependent inhibitory effect on the RRDA, prolonging the respiratory cycle and expiratory time and decreasing the integral amplitude and inspiratory time. EGLU induced a significant decrease in the respiratory cycle and expiratory time. The effect of APDC on the respiratory rhythm was partially reversed by the application of APDC+EGLU.

CONCLUSIONGroup II metabotropic glutamate receptors are probably involved in the modulation of the RRDA in isolated neonatal rat brainstem slice.

Animals ; Animals, Newborn ; In Vitro Techniques ; Medulla Oblongata ; physiology ; Rats ; Rats, Sprague-Dawley ; Receptors, Metabotropic Glutamate ; physiology ; Respiratory Center ; physiology

OBJECTIVETo investigate the effects of doxapram on the respiratory rhythmical discharge activity (RRDA) in the brainstem slices of neonatal rats.

METHODSThirty neonatal SD rats (of either sex, 0-3 days old) were randomly divided into 6 equal groups (groups I-VI), and the brainstem slices which contained the medial region of the nucleus retrofacialis (mNRF) were prepared. All the slices were perfused with modified Kreb's solution (MKS), and in group I (control group), the slices were perfused with MKS only; in groups II to IV, the slices were perfused with doxapram in MKS continuously at the concentrations of 2, 5, and 10 micromol/L, respectively; in groups V and VI, the slices were perfused with 20 micromol/L propofol and 20 micromol/L propofol plus 5 micromol/L doxapram, respectively. The RRDA in the hypoglossal nerve was recorded by suction electrode. The discharge time course of the inspiratory (TI), expiratory (TE), respiratory cycle (RC) and integral amplitude of the inspiratory discharge (IA) were recorded at 1, 3, 5, 10, 15, and 30 min after the application of the drugs.

RESULTSThe hypoglossal nerve in groups I, II and VI showed no significant changes of RRDA in the entire course of the experiment (P>0.05). In groups III and IV, the TI, IA increased and TE decreased significantly 5 min after doxapram application (P<0.05), and the RC was shortened only at 10 min. In group V, the TI and IA decreased and the RC and TE increased significantly after the drug application (P<0.05).

CONCLUSIONDoxapram (>5 micromol/L ) can directly stimulate the RRDA and prevent propofol-induced inhibitory effects in the brainstem slice of neonatal rats, and the effects are mediated by its actions upon the inspiratory neurons in the mNRF.

Animals ; Animals, Newborn ; Doxapram ; pharmacology ; Electrophysiological Phenomena ; drug effects ; Female ; In Vitro Techniques ; Male ; Medulla Oblongata ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Respiration ; drug effects ; Respiratory System Agents ; pharmacology

Objective To study the bone biomechanics of the rabbit osteoporosis models induced by dexamethasone sodium phosphate injection (DX) using a combined treatment modality of 99Tc-MDP and GuKangLing.Methods Rabbits were intramuscularly injected with DX (2 mg/kg) twice a week for 6 weeks.The animal osteoporosis model group (Group C) and normal group (Group A) were compared to confirm the model was available.Another control group (Group B),the osteoporosis control group (Group D) were set for the comparison at the end of the experiment.The 99Tc-MDP therapy group (Group E),GuKangLing therapy group (Group F) and 99Tc-MDP plus GuKangLing therapy group (Group G) were included in the study.The treatment lasted for 16 weeks.The bone biomechanics,cytopathology bone histomorphology,bone mineral density (BMD),X-ray,CT,bone scintigraphy and serum bone alkaline phosphatase (BALP)and P (bone gla protein) were chosen as the markers or methods to evaluate the treatment results (excellent,effective and invalid).The analysis of variance (ANOVA) and t-test were used for group comparison analysis.Results Cytopathology result indicated that there was no bone trabecula destruction in Group A.However,there was distinct bone destruction in Group C.The bone biomechanics (left femur head (265.914 ±52.773) N,L4(369.671 ±94.919) N),BMD(left femur (0.238 ±0.016) g/cm2,L4(0.236 ±0.016) g/cm2)and bone histomorphology ( (66.230 ± 10.848) ％ ) in Group C reduced clearly as compared with Group A ((405.343±55.410) N,(750.870±53.718) N,(0.294±0.017) g/cm2,(0.302±0.023) g/cm2,( 131.500 ± 21.846) ％ ) ( t ≥4.550,all P ＜ 0.01 ).Radionuclide bone scan also showed that the uptake of tracers was higher by the main arthrosis in Group C than that in Group A.Vertebra was not clearly visualized on bone scan image.There were significant differences between Group A and Group C in serum BALP and P ((45.000±7.303) vs (12.485 ±1.512) U/L,(0.168±0.018) vs (0.115 ±0.017) μg/L,t =4.126,5.476,both P ＜ 0.01 ),which indicated that the animal osteoporosis model was available.The pathological results showed an improved recovery of bone structure and trabecular in Groups E and G,but a worse recovery in Group F.Biomechanics result in Groups E and G (left femur head (386.457 ±77.077) N and (432.771 ± 17.525) N,L4(649.550 ± 126.859) N and (655.443 ±76.555) N) improved apparently,which were similar to Group B.The radiotracer uptake in Group F was lower than that in group D.The bone biomechanics,bone histomorphology,BMD,serum BALP and P after the treatment showed significant differences in Groups E,F and G (F:8.556 - 31.608,all P＜0.01 ),and the bone biomechanics result in Group G was a little better than that in Group E (t =2.625,P ＜ 0.05 ).The results of Group G and E were considered as excellent,and Group F was considered as effective.Conclusions The treatment of 99Tc-MDP combined with GuKangLing could improve the bone biomechanics of rabbit osteoporosis models and may be a potential method to increase the bone strength for resisting external force.

Objective To explore the best operation time and approach, and the improvement mechanism of radiation encephalopathy in patients with nasopharyngeal carcinoma. Methods The data of 9 patients with radiation encephalopathy after radiotherapy for nasopharyngeal carcinoma who admitted to our hospital from January 1996 and December 2008 were retrospectively analyzed. The neurological manifestations, imaging, neurosurgery strategies and pathological features of the patients were collected and analyzed. The efficacy and the follow-up results were compared. Results MRI showed such typical encephalopathy as severe edema or necrosis in the temporal lobe in most patients adopted radiotherapy; edema and necrosis in the brain tissues, hyaline degeneration and blocking in the small blood vessels were showed by pathologic examination. Some patients manifested as having unilateral edema or liquefactive necrosis in the brain with marked mass effect. After the surgery, the edema in the patients' brain was alleviated; 8 patients got improvement and discharged from the hospital in 2-3 weeks with stable vital signs. Eight patients achieved complete remission of headache with 1 having mild headache. Conclusion When medical treatment is not effective in patients who developed edema or liquefactive necrosis after radiotherapy, neurosurgery is a good therapeutic strategy, which can alleviate the symptoms, help clarify the diagnosis and guide the follow-up treatment.