Objective To investigate the tumor-inhibitory effect of cytokine-induced killer cells(CIK)co-cul-tured with dendritic cells (DC)pulsed by breast cancer stem cell antigen on the same tumor-bearing mice. Methods Breast cancer stem cells were isolated from the cell line of MCF-7/ADR and extract lyses antigen of the stem cell was saved. DC and CIK derived from peripheral blood mononuclear cells of healthy individuals were co-cultured and pulsed or un-pulsed by the above antigen lyses. This DC+CIK were injected to breast tumorbearing mice (BCSC-AP-DC+CIK group), and were used to compared with the common breast cancer cell antigen (rather than breast cancer stem cell antigen) pulsed DC+CIK group(AP-DC+CIK group), DC+CIK group, CIK CIK group and normal saline group(NS group). The tumor-inhibitory effect were evaluated and compared among all 5 groups through the tumor size, TdT-mediated dUTP nick end labeling test (TUNEL), examining expression level of bcl-2 and bax by immunohistochemistry. Results The tumor size in each group before and after therapy and the tumor size after therapy between each group was of significant difference(P＜0.05). The maximum size is NS group(3.625±0.093)cm3 and BCSC-AP-DC+CIK group is minimum,which is (1.234±0.131)cm3. BC-SC-AP-DC+CIK group is of highest expression of bax and apoptotic index value, lowest bcl-2 expression in all 5 groups. Conclusion The CIK co-cultured with DC pulsed breast cancer stem cell antigen was more effective to induce apoptosis of breast cancer cells than those of CIK cells co-cultured with DC pulsed breast cancer cell antigen,CIK cells co-cultured with DC and CIK cells.

Background Optic neuritis is one of the common clinical neuro-ophthalmic diseases.Recurrent optic neuritis can cause irreversible axonal damage and visual impairment,and no prevention method is effective up to now.Its pathogenesis is considered to be associated with the body's immune imbalance.Objective This proposal tried to establish experimental autoimmune encephalomyelitis (EAE) and explore the immunomodulatory effects of T helper (Treg),Th17 cell in the model of optic neuritis.Make the immune mechanism of demyelinating optic neuritis clear.Methods EAE model was established to observe the clinical symptoms and visual electrophysiological changes of optical neuritis in mice;hematoxylin-eosin staining to observe the histological changes in the mouse models of EAE; immunohistochemical staining to detect optic nerve axon damage markers β-amyloid peptide precurso (β-APP) protein expression quantity changes;reverse transcription PCR (RT-PCR) to detect the dynamic expression of forkhead/winged helix transcription factor p3 (Foxp3),transforming growth factor-β (TGF-β),interleukin-1β (IL-1β),IL-17,IL-10 in optic nerve tissue.Results After immunization 11 days,neurological symptoms of EAE mice started to appear,hematoxylin-eosin staining showed optic nerve tissue infiltration of inflammatory cytokines; immunohistochemical staining showed enhancement of axonal injury markers β-APP positive staining; flash visual evoked potential(F-VEP) test showed 7,11,14,19,23,28 days,compared with the normal mice,P1 prepatents of model group were prolonged after modeling,the differences were statistically significant (t =4.487,15.203,16.364,11.540,11.959,16.163,all at P＜0.05).When 7 days after modeling,the difference of N1-P1 amplitude between normal group and model group was no statistical significance (t =-0.992,P =0.378); while 11,14,19,23,28 days after modeling,compared with normal mice,N1-P1 amplitudes of model mice were significantly lower,the differences were statistically significant(all at P＜ 0.05).The expressions of TGF-β,IL-1β,IL-17,Foxp3,IL-10 mRNA among different time points had stastistie significant differences (F =12.721,15.015,14.343,69.374,68.290,all at P =0.000),compared with the normal group,when 11 days,14 days after modeling,TGF-β mRNA levels were significantly increased ; 19 days,23 days after modeling,IL-1β mRNA levels were significantly increased;7 days,11 days after modeling,IL-17 mRNA levels were significantly increased ;7,11,14,19,23,28 days after modeling,Foxp3 mRNA levels were significantly lower; 19,23,28 days after modeling,IL-10 mRNA levels were significantly lower,the difference were statistically significant (all at P＜ 0.05).Conclusions Th17 cells launch demyelinating optic neuritis immune injury,and Th1 subgroup performs the function of maintaining inflammatory lesions,Treg cell subgroup is abnormal much earlier than that of the Th2 subgroup.The imbalance of Th17/Treg cell may be involved in the pathogenesis of demyelinating optic neuritis.

Background Trans-corneally subretinal injection in rodent model is a useful method for genetic therapy,stem cell transplantation and the study on the ophthalmic research.Standarized operation process is critical for the successful treatment.However,there is no literature to report the detailed procedure and the influence of this technique on morphology and function of retina.Objective This sudy was to introduce a method of trans-corneally subretinal injection and evaluate its influence on the morphology and function of retina.Methods Trans-corneallly subretinal injection was performed on the left eyes of 2-month-old SPF C57BL/6J mice after dilation of pupils.A 301/2G disposable needle was used to puncture the cornea within the pupil area near limbus and avoid touching the lens and irises under eye surgery microscope.Then,a 33G blunt needle was used to insert into the vitreous and toward subretinal space via corneal puncture.Normal saline with 0.1％ fluorescein sodium of 1 μl was slowly injected into the space,and 2.5％ hydroxypropyl methylcellulose was dropped on ocular surface for the observation of the fundus clearly.According to the percentage of the retina filled with subretinally injected solution,the experimental eyes were divided into 80％-100％ area group,50％-70％ area group after injection,and the mice in the pseudo-injected group,in which injection procedure stopped just before the solution was pushed in to the subretinal space did not inject any solution after punctured.The right uninjected eyes of the mice served as normal control group.Four eyes were selected for each group.The structural changes were evaluated by optical coherance tomograpby (OCT) 1 day,2 days,3 days and 5 weeks after injection,and retinal function was assessed by the recored of electroretinography (ERG) 5 weeks after injection.The retinal sepcimens were prepared to examin the morphological changes by hematoxylin and esosin staning.The use of care followd the Regulations for the Administration of Affair Concerning Experimental Animals of Zhejiang Province.Results About 70％ of the injected eyes showed that retinal blebs filled with injected green fluorescein solution occupied 50％ or more retinal area with minimal damages.The focal detachment between neurosensory retinal layer and retinal pigment epithelium (RPE) was exhibited 1 day postinjection,and almost all the retinas retached 2 days after injection.In the fifth week after injection,the amplitudes of ERG b wave were (386.25±37.88),(357.50±41.03),(324.25±53.45) and (410.50±14.88) μV in the sham operation group,50％-70％ area group,80％-100％ area group and normal control group,respectively,showing a significant difference among the 4 groups (F=3.574,P=0.047),and the amplitudes of b wave in the normal control group were higher than those in the 80％-100％ area group (all at P ＜ 0.05).The detachment between retinal neuroepithelium layer and RPE layer,cell proliferation and transposition in the outer nuclear layer were dispalyed under the light microscope in the sham operation group,50％-70％ area group and 80％-100％ area group,and the disordered outer segment of photoreceptors at the injecting area was seen in the 50％-70％ area and 80％-100％ area groups at five weeks after injection.However,retinal sructure and morphology were normal at the non-injection area.Conclusions Trans-corneally subretinal injection is an effective and safe way for subretinal injection.

Background The retinal degeneration 11 (rd11) mouse is a newly discovered naturally occurring recessive animal model with lysophosphatidylcholine acyltransferase 1 (Lpcatl) mutation.Previous studies showed that the photoreceptor cells are characterized by typical rod-cone degeneration pattern in rd1 1 mice,while cone degeneration pattern in rd11 mice is unclcar.Objective Using immunofluorescence staining techniques with retinal wholemount,we aim to clarify the degeneration patterns of cone-function related M-opsin or S-opsin in different ages of rd1 1 mice.Methods A total of thirty rd1 1 and C57BL/6J mice at postnatal (P) day 14,28,42 (five in each age group) were sacrificed and retinal wholemounts were prepared.Immunohistochemistry was performed to identify the expression of M-opsin or S-opsin in retinal wholemounts,which were photographed with a fluorescent microscope.Cone opsins were compared between rd1 1 retinas and age-matched normal C57BL/6J retinas by manually counting the opsin positive cone cells in different quadrants of the retinas.Results The number of M-opsin or S-opsin positive fluorescent dots in each quadrant was similar at all ages of normal C57BL/6J retina.M-opsin positive fluorescent dots in dorsal/temporal,ventral/temporal,dorsal/nasal and ventral/nasal quadrants of rdl 1 retina at P28 were (414±32),(300± 8),(324 ± 22) and (250± 20)/0.037 mm2,which were lower than the age-matched normal C57BL/6J mice (t =4.114,15.225,7.505,17.990,all at P＜0.05).At the same time the S-opsin positive fluorescent dots in P28 rd11 were (8 ±4),(175 ± 16),(74 ± 13) and (315 ±20)/0.037 mm2,with significant decrease in comparison with those in the age-matched normal C57BL/6J mice (t =8.555,17.076,21.637,13.498,all at P＜0.05).With the development of retinal degeneration in rd11 mice,the M-opsin degeneration spread from central to ventral,nasal and then to temporal and dorsal peripheral retina;and the S-opsin loss started from dorsal/temporal to ventral/nasal retina.Conclusions Most of the M-opsin and S-opsins,especially the S-opsins in rd11 mice,degenerate in 6 weeks.Retinal wholemount and cone opsin immunofluorescent staining provide a useful tool to show the cone degeneration pattern and to evaluate the therapeutic efficiency in ongoing gene therapy study.

Anticonvulsants ; therapeutic use ; Child ; Child, Preschool ; Electroencephalography ; Epilepsies, Partial ; diagnosis ; drug therapy ; Humans ; Infant ; Occipital Lobe ; pathology ; Prognosis

AIM: To measure the macular thickness of normal young people by 3D 1000 optical coherence tomography (OCT) and study the repeatability of measuring results and the relationship between the thicknesses of macular and gender. At the same time, to compare our result with the data of other types of OCT, and to understand the consistency of the measuring results of macular thickness of different types of OCT.
METHODS: Totally 222 eyes in 111 young people were detected using 3D scan mode of Topcon 3D OCT 1000 (ver 2.4 ) . Twelve cases ( 24 eyes ) underwent repeatability check. We took transverse comparison between our measured results with other research's results.
RESULTS: There were 111 cases of young people, whose age were from 18-27 years old, all uncorrected and corrected visual acuity were≥1. 0, all intraocular pressure were <21mmHg. The average thickness of all macular region was 273. 32±17.08μm. Retinal volume of macular area was 7. 73 ± 0.37mm3 . Center thickness was 161 -264μm, and the average thickness was 200. 13±18. 81μm. Central macular thickness were 188 - 273μm, and the average thickness was 229. 00 ± 18. 20μm. The central macular thickness in men was significantly greater than that in women, and there was statistical difference. The results of repeated check of 12 cases ( 24 eyes ) in the macular area were no statistical difference except the outer ring of nasal quadrant, and the repeatability of average thickness in central macular thickness was better than in center thickness.
CONCLUSION:The repeatability of macular examination is good. The central macular thickness can be better repeated than the center thickness. The central macular thickness is 229. 00±18. 20μm in young people, according to the 3D 1000 OCT measurements. There are statistical difference of central macular thickness between different genders.

Objective To compare different beginning time of acid simulation to reduce the acute damage of salivary glands after high-dose of iodine-131 treatment for the post-surgery patient with differentiated thyroid cancer (DTC) and screen out the optimal beginning time of acid simulation .Methods Total 309 cases of post-surgery DTC patients accept high-does ioding-131 treatment (average dose of ioding-131 :4 .28 ± 0 .43 GBq) ,and all patients was divided into three group randomly ,the beginning time of acid stimulation(vitamin C :10 mg per time ,three time a day ,lemonade:50 mL per 2 h) for each group is 2 ,12 and 24 h .Then observe the incidence and time of acute damage of salivary glands for each group .At same time ,we analyses the relation between acute dam-age of salivary glands with sex and age .Results The incidence of acute damage of salivary glands is 13 .21% (2 h) ,24 .51% (12 h) , 26 .73% (24 h)respectively ,the incidence of 2 h is lower than those of 12 h and 24 h obviously(P<0 .05) ,and there is no difference between 12 h and 24 h(P>0 .05) .The occurrence time of acute damage of salivary glands for 84 .85% patients is between 10-24 h .And there is no difference of incidence of acute damage of salivary glands between different sex and age group .Conclusion 2 h maybe the optimal beginning time of acid simulation to reduce the acute damage salivary glands for the post-surgery DTC patients after high-dose iodine-131 treatment in this study .Sex and age are no influence to the occurrence of acute damage of salivary glands .

Objective To explore the relationship between cardiac involvement and laboratory indicators in patients with rheumatoid arthritis(RA). Methods Cardiac echocardiography and ECG performance of 184 patients with RA were analyzed retrospectively. Results Among the 184 patients with RA, the pulmonary hypertension detection rate was 8. 3%, valvular disease 38. 9%, arteriosclerosis 27. 8%, wall to reduce the exercise 13.9%, myocarditis 5.6% and pericardial effusion 5.6%, according to the echocardiography examinations;Sinus tachycardia was evidenced in 15. 22% patients, ST-T changes in 39. 13%, electric axis left side in 8. 70%, branch block in 13.04%, left ventricular hypertrophy in 4. 35%, atrial fibrillation in 4. 35%, premature in 8.70%, early repolarization syndrome in 2. 17% and electric-axis right side in 4. 35% patients by ECG examinations. The serum level of CRP (46. 77 ±5. 87) mg/L was significantly higher in RA patients with cardiac involvement than that in the non-cardiac involvement patientsm (28. 45 ±3. 21) mg/L (P ＜0.05) ;While the serum level of ESR,RF,IgG,IgA,IgM, PLT showed no statistically significant differences between the two groups (P ＞ 0.05); Within RA patients withcardiac involvement, the serum level of CRP showed no significant difference among different sub-groups , which were classified according to the echocardiography performance (P ＞ 0.05). Conclusions Cardiac involvement occurred frequently in patients with rheumatoid arthritis. The valvular disease, arteriosclerosis, reducing of the wall motion and pericardial effusion are the main manifestations by echocardiography examination; Sinus tachycardia, ST-T changes,branch block and premature beats are the main ECG abnormalities. The serum level of CRP is significantly higher in RA patients with cardiac involvement than that with non-cardiac involvement patients. The higher level of CRP in patients with RA may indicate the cardiac involvement presence.

Objective To investigate the pathogenesis of the expression and activation of signal transducers and activator of transcription(STAT) 6 in patients with ulcerative colitis(UC).Methods Thirty patients with active UC(diagnosed with colonoscopy with pathologicaly conformation),who were not treated with steroids or immunomodulators,and 30 age-matched healthy controls were studied.The expressions of phosphorylated STAT6 and non-phosphorylated STAT6 in nucleus and cytoplasm were deterimined by Western blot.DNA binding activity of STAT6 was tested by electrophoresis mobility shift assay(EMSA).Results The expression of phosphorylated STAT6 was higher in nucleus than that in cytoplasm in mild UC patients,but the expression was higher in cytoplasm than that in nucleus in moderate to sever UC patients (P

Objective To evaluate the results of hybrid procedure in treating infants and children with severe pulmonary stenosis (PS) and the clinic value of echocardiography in the whole procedure.Methods Hybrid therapy was performed in 43 patients [(7.1±6.8)months,ranging 5 days to 33 months] with severe PS.The echocardiography data in the whole procedure and follow up were reviewed.Results Aided by echocardiography,the hybrid therapy was successful in 43 cases without major complication.Immediately following valvuloplasty,the gradient across pulmonary valve decreased from (92.5±21.4)mm Hg to (23.6±13.0)mm Hg (P<0.05).The growth and development of 27 patients in follow-up study was well.No restenosis was found and tricuspid regurgitation decreased differently.Conclusions Hybrid procedure is effective and safe for the treatment of infants and children with severe PS.And echocardiography play an important role in the whole procedure.