Objective To investigate whether RelB-silenced bone marrow-derived dendritic cells (BMDC) pulsed with torpedo acetylcholine receptor (TAChR) immuno-dominant peptide Tα_(146～162) can induce tolerance in T cells primed with TAChR. Methods Recombinant lentivirus that produced RelB siRNA and control lentivirus were prepared and used to infect BMDCs. The infected BMDCs were stimulated with LPS,and the resulting cells were designated as DC-siRelB or DC-control, respectively. The mRNA and protein expression of RelB were examined by quantitative real-time PCR and Western blot. Cell surface markers of DC were evaluated by flow cytometry. IL-12 in the supernatant was detected by ELISA. Mice were randomly divided into 6 groups: A1, A2, A3,K1, K2, and K3. On day 0, group A1, A2, and A3 were primed with TAChR in CFA and group K1, K2 and K3 were primed with KLH+CFA. On day 7, group A2 and K2 were injected with Tα_(146～162) pulsed DC-siRelB, group A3 and K3 were injected with Tα_(146～162) pulsed DC-control, while A1 and K1 group received PBS at the same time. On day 14, lymphocyte proliferative response of the 4 groups were measured. Results Recombinant lentivirus including RelBshRNA genes was successfully constructed. RelB siRNA knocked down RelB expression in BMDCs obviously. Compared with DC-control, DC-siRelB expressed a significantly lower level of CD80, CD86, and MHC class II on their surface, producing lower level of IL-12. Compared with group A1 and A3, lymphocyte proliferative response to TAChR of A2 group was suppressed significantly (P<0.05). No different lymphocyte proliferative responses to KLH and ConA were seen in group A1, A2 and A3 (P>0.05). No different lymphocyte proliferative responses were seen in group K1, K2 and K3 (P>0.05). Conclusion Lentiviral-mediated RelB-silenced BMDCs are maturation resistant and can induce antigen-specific tolerance in TAChR primed C57BL/6 mice,which provides a basis for further study of their therapeutic potential in myasthenia gravis.

Objective To evaluate the anatomic changes and dosimetric variations of patients with nasopharyngeal carcinoma (NPC) during the course of simultaneous integrated boost intensity-modulated radiotherapy (SIB-IMRT) by comparison of the dosimetric differences with or without replanning.Methods Twelve cases with NPC treated with SIB-IMRT underwent repeated CT scans after 20- 25 fractions of the initiation of therapy.The original treatment plan ( Plan1 ) based on the first CT scan ( CT1 ) and the second IMRT plan (Plan 2) based on the second CT scan (CT2) were calculated with an inverse planning system (Pinnacle3,Philips Medical System).In addition,the hybrid IMRT plan,Planl (CT2),was generated by deformable registration with MIMVISTA software,and the doses in Plan 1 ( CT1 ) and Plan 2 ( CT2 ) were accumulated based on CT2.The dosimetric differences were compared among the Plan 1 ( CT1 ),Plan 1 (CT2) and Plan 1 + 2(CT2).Results Compared with CT1,the mean volumes of the right and left parotid glands in the CT2 were significantly smaller by ( 24.6 ± 11.9 ) ％ and ( 35.1 ± 20.1 ) ％,respectively.Compared with Plan 1 ( CT1 ),the dose received by 95％ of the target ( D9s ) to PGTV,PTV1 and PTV2,and mean dose (D ) to PGTV,and PTV2 were all significantly lower in the Plan 1 (CT2),indicating that the doses to targets decreased without replanning.With repeated CT and replanning after 25 fractions as shown in Plan 1 + 2 (CT2),the doses to targets would be improved.The doses to normal tissue were increased without replanning,although no statistical significance was observed.In 5 of 12 cases,the doses to the spinal cord and brainstem exceeded the constraint without replanning,while the corresponding values decreased with replanning.Conclusions During the course of IMRT for cases with NPC,the volumes of the targets and parotid glands decrease significantly.Mid-treatment CT scanning and replanning should be recommended to ensure adequate doses to the targets and safe doses to the normal tissues.

Objective To obtain colon cancer spheroid cells from human colon cancer cell lines cultured in serum-free medium (SFM),and investigate the proliferative and migratory properties of colon cancer spheroid cells.Methods Human colon cancer cell lines HCT116 and HT29 were cultured in SFM,and then the generation of spheroid cells was observed.The expression of stem cell surface marker CD133 was detected by flow cytometry,and the proliferative and migratory properties of colon cancer spheroid cells were detected by cell counting kit-8 and Transwell migration assay,respectively.All data were analyzed by using the t test.Results Spheroid cells were obtained from colon cancer cell lines HCT116 and HT29 in SFM.The ratios of spheroid cells with positive expression of CD133 generated by HCT116 and HT29 were 75.44％ ± 11.41％ and 76.22％ ± 14.23％,respectively.Compared with original colon cancer cells cultured in serum supplemented medium,the number of HCT116 and HT29 spheroid cells with positive expression of CD133 was significantly greater (t =11.43,9.17,P ＜ 0.05 ),and the proliferative and migratory abilities were much stronger also.Conclusion Colon cancer spheroid cells cultured in SFM have higher positive expression of CD133 and stronger proliferative and migratory abilities,and it can be utilized as a feasible model for further studies of colonic stem cells.

Objective To evaluate the dose distribution of target volume and normal tissues in forward intensity modulated radiotherapy (fIMRT) and inverse intensity modulated radiotherapy (iIMRT) modes for breast cancer after radical mastectomy.Methods Both fIMRT and iIMRT plans were developed for 10 patients with breast cancer after radical mastectomy.On each patient's CT images the supraclavicular area, chest wall, and internal mammary area were delineated.The prescription dose was 50 Gyin 25fractions.In the fIMRT plan X-ray irradiation at the dose of 6 MV was adopted for the supraclavicular and the chest wall areas and electron irradiation at the dose of 9 - 12 MeV was adopted for the internal mammary area, and the doses of cold and hot spots were adjusted according to the fitting doses of these 3 regions.In the iIMRT plan the supraclavicular area, chest wall, and internal mammary area were taken asa whole target, 6 MV X-rays was used, and inverse optimal design was performed.The dose distribution oftarget volume and normal tissues, conformal index (CI) , and heterogeneous index (HI) , and acceleratormonitor unit (MU) were analyzed using dose-volume histogram (DVH)for the two intensity modulated modes.Results The maximum dose of PTV of the iIMRT plan was significantly lower than that of the fIMRT plan(t = -3.23,P ＜0.05), the minimum dose and V95% of PTV of the iIMRT were significantly higher than those of the fIMRT plan(t = 4.08, -2.69, both P ＜0.05).The CI level of the iIMRT plan was significantly higher than that of the fIMRT plan and the HI level of the iIMRT plan was significantly lower than that of the fIMRT plan (t = -3.13, 2.74, both P ＜0.05).There were not significant differences in V10, V20, V25, V30, and Dmean of the ipsilateral lung between these 2 groups.However, the V15 of ipsilateral lung of the iIMRT group was significantly lower by 4.2% than that of the fIMRT group (t= 3.2, P ＜ 0.05).There were not significant differences in the mean dose (Dmean) and V30 of heart, and Dmean of contralateral lung and contralateral breast between these 2 groups.Conclusions Compared with fIMRT, the iIMRT plan results in more PTV coverage, higher conformity index, and more homogeneous dose distribution, with lower dose upon the lung at the affected side, and better protection of the contralateral lung, heart, and breast.

Administration, Inhalation ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Methemoglobin ; analysis ; Nitric Oxide ; administration & dosage

OBJECTIVE To investigate the molecular mechanisms of resveratrol( Res)in renal interstitial fibrosis(RlF)in rats with unilateral ureteral obstruction(UUO). METHODS Forty-eight Spra-gur-Dawley rats were randomly divided into UUO( normal saline,n = 16),UUO with Res treatment (Res,20 mg·kg-1 ,n=16),and sham-operation(sham,n=16)models. The kidneys were excised on the 7th and 14th day. The deposition of collagen fiber in the kidney was detected with HE and Masson staining. The levels of sonic hedgehog(SHH,an inducer of SHH pathway)in kidney tissues were deter-mined by ELlSA. lmmunohistochemical analysis was performed to evaluate the protein expression of SHH signaling-related molecules,including SHH,smoothened(Smo),patched-1(Ptch1),and Gli1, proliferating cell nuclear antigen(PCNA)and matrix component typeⅢ collagen. The mRNA expression levels of Smo,Ptch1 and Gli1 were detected by real-time RT-PCR. RESULTS The degree of RlF observed with HE and Masson staining was obviously increased in UUO kidneys,but decreased in Res-treated kidneys. Enhanced expression levels of typeⅢ collagen and PCNA in UUO rats were suppressed by Res treatment(P﹤0.05). Res administration decreased the expression levels of SHH,Smo,and Gli1 (P﹤0.05),but increased the expression of Ptch1(P﹤0.05),suggesting that Res inhibit the obstruction-induced activation of SHH signaling. CONCLUSION Res can attenuate RlF in UUO rats,and the possi-ble mechanism is that Res down-regulates the activity of SHH signaling and inhibits cellular proliferation, resulting in inhibition of matrix accumulation in renal interstitium of UUO rats.

Objective To analyze the characteristics of bone scintigraphy in 117 cases with primary hyperparathyroidism (PHPT).Methods Of these 117 cases (50 males and 67 females),there were 116 parathyroid adenomas and 1 parathyroid cancer.Mean age was 61.1(12-86) years old.All had ~(99)Tc~m-methylene diphosphonate (MDP) bone scintigraphy.The bone images could be classified into 4 categories.Category Ⅰ:normal;category Ⅱ:localized abnormal,which could be subcategorized as Ⅱ A with skull and mandible involvement,and Ⅱ B with Ⅱ A characteristics plus metabolic derangement;category Ⅲ:systemic,whole-body incmased tracer uptake;category Ⅳ:systemic plus localized metabolic derangement.Data were analyzed statistically with X~2 and t-test (isolated samples).Results According to the scintigraphic findings,there were 47 cases(40.17%)of category Ⅰ,35 cases(29.91%) category Ⅱ (21/35cases Ⅱ A and 14/35 cases Ⅱ B),30 cases (25.64%) category Ⅲ,and 5 cases (4.27%) category Ⅳ.Combining categories Ⅱ、Ⅲ and Ⅳ together,there were 70 abnormal cases.These patients had history of abnormal bone images such as bone fracture (39 cases,55.71%),calculus (8 cases,11.43%),bone fracture plus calculus(7 cases,10.00%),osteoporosis (51 cases,72.86%) or ostalgia(26 cases,37.14%);however,in the 47 cases of category Ⅰ,only 1 (2.13%),0,0,10(21.28%)and 10 cases (21.28%),respectively,were found.Therefore.these case history characteristics were statistically significant (X~2=11.152,P=0.01).The tumor size,parathyroid hormone (PTH),blood calcium,blood phosphorus in the patients of abnormal PHPT categories Ⅱ to Ⅲ were(14.52±13.72)cm~3,(731.67±618.40)ng/L,(3.05±0.29) mmol/L and (0.71±0.14) mmol/L,respectively.with statistically significant difference compared to category Ⅰ:(0.78±1.33) cm~3,(112.04±62.98)ng/L,(2.56±0.42) mmol/L and (1.03±0.36)mmol/L(t=-5.724,-5.741,-7.274 and -6.451;all P＜0.01).Conclusions (1)Bone scintigraphy was normal in 40% of PHPT patients.(2)The bone images of PHPT could be classified into 4 categories and each could reflect the duration and severity of the disease status on bone.(3)The bone imaging characteristic could be useful for differential diagnostic purposes.

Objective To analyze the characteristic of trauma in 246 fatal falls. Methods We collect 246 cases of fatal falls from Pudong district of Shanghai from 2005 to 2014, and then analyze the characteristics of trauma by using different parameters. Results The height in suicides by fall is higher than accidents by fall (25.7m:13.98m). In all fatal falls, head has a higher possibility in suffering trauma than other body regions (75.2%), while neck has a lower possibility (6.5%). The possibility of head trauma in accidents by fall is higher than suicides by fall (89.4%:75.2%). Conclusion Comparing with suicides by fall, victims in accidents by fall have lack of protection from limbs. Besides, the low rate of ifnding neck trauma suggests legal physicians should be more careful in examining this area.

Objective To understand the prevalence,drug resistance and clinical features of children with acute lower respiratory tract infection(ALRI) caused by escherichia coli.Methods From Oct. 2005 to Oct. 2006,659 patients with ALRI who were admitted to hospital were chosen and their nasopharyngeal secretions were obtained and cultured.K-B disc diffusion for antibiotic susceptibility were performed for these clinical isolates.Results Among 659 patients,118 cases were isolated escherichia coli,the rate was 17.99% which had 90 boys and 28 girls.Eighty-seven of 118 E.coli strains were with extended-spectrum ?-lactamase(ESBLs),the rate was 73.73%.All of strains were sensitive to imipenem.For ESBLs-producing strains,the ratio of resistance tocefotaxime,ceftriaxone,cefuroxime,ampicillin,piperacillin were 78.81%,73.73%,73.73%,76.27%,78.81%,respectively.Conclusions The positive rate of ESBLs producing E coli in Kunming area is high and drug resistance is severe gradually.Imipenem can be the first selection for treatment on these infections.

Along with the development of economy and society, type 2 diabetic mellitus (T2DM) has become one of the most common diseases at the global level. As one of the complications of T2DM, diabetic neuropathic pain (DNP) stubbornly and chronically affects the health and life of human beings. In the pain field, dorsal root ganglion (DRG) is generally considered as the first stage of the sensory pathway where the hyperexcitability of injured neurons is associated with different kinds of peripheral neuropathic pains. The abnormal electrophysiology is mainly due to the changed properties of voltage-gated sodium channels (VGSCs) and the increased sodium currents (I(Na)). Curcumin is an active ingredient extracted from turmeric and has been demonstrated to ameliorate T2DM and its various complications including DNP effectively. The present study demonstrates that the I(Na) of small-sized DRG neurons are significantly increased with the abnormal electrophysiological characteristics of VGSCs in type 2 diabetic neuropathic pain rats. And these abnormalities can be ameliorated efficaciously by a period of treatment with curcumin.
Animals ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; complications ; Diabetic Neuropathies ; drug therapy ; Ganglia, Spinal ; cytology ; drug effects ; metabolism ; Neuralgia ; drug therapy ; Neurons ; drug effects ; metabolism ; Rats ; Sodium ; Voltage-Gated Sodium Channels ; physiology