Background Cell immunologic therapy for retinoblastoma(RB)is becoming a hot research topic.Cancer-testis antigen is a human immunogenic protein and is used to treat some tumors.However,its effect on RB has not been investigated.Objective The present study was to discuss the antigen specific anti-tumor effect of cytotoxic T lymphocytes(CTL)induced by the cancer-testis antigen,NY-ESO-1-sensitized dendritic cells(DCs),on human RB.Methods PCR was performed to amplify target gene fragments from the NY-ESO-1 plasmids,and then the target gene fragments were digested with the restriction enzymes SalI and EcoRI.Harvested fragments were inserted into the pDC316 plasmid to construct the recombinant plasmid pDC316/NY-ESO-1.The expression of NY-ESO-1 protein in human RB cells strain,HXO-RB44,was detected by immunofluorescence and Western blot.Monocytes were isolated from 60 ml of peripheral blood from a healthy donor using Ficoll density-gradient centrifugation with a cell density of 1 × 107/ml.DCs isolated from blood were stimulated with recombinant human granulocyte-macrophage colony stimulating factor(rhGM-CSF)and recombinant human interleukin-4(rhIL-4).The recombinant plasmid pDC316/NY-ESO-1 was transfected into DCs and the DCs were co-cultured with T lymphocytes.The resultant CTL were used as effector cells.The growth of the CTL was detected by MTT assay.The CTL were then added into the growth medium used for culturing HXO-RB44 cells and the vitality of the HXO-RB44 cells was assayed by MTT assay.Results The sequence of the cloned DNA fragment of the recombinant plasmid pDC316/NY-ESO-1 was conforms with the sequence of the NY-ESO-1 gene.The expression of the NY-ESO-1 protein in HXO-RB44 cells was tested by immunofluorescence and Western blot.DCs were successfully induced with rhIL-4 and rhGM-CSF from PBMC.The recombinant expression plasmid pDC316/NY-ESO-1 was successfully transferred into DCs.These DCs had high expression of surface molecules such as HLA-DR(42.1％),CD80(54.2％),CD83(39.7％)and CD86 (94.8％).The CTL that was induced by DCs-sensitized with NY-ESO-1 specifically killed HXO-RB44 cells.CTL induced by the sensitized DCs had a stronger cytotoxic effect against HXO-RB44 cells compared with un-sensitized DCs and CTL un-induced with DCs,as shown by MTT asssay(P＜0.05).The anti-tumor activity was highest when the ratio of effector to target was 75∶1(P＜0.05).Conclusions DCs transfected by the recombinant plasmid pDC316/NY-ESO-1 can induce the proliferation of allogenic CTLs,which showed a specific anti-tumor effect against HXO-RB44 cells.These results present a new type of immunotherapy for the treatment of RB.

Bone Diseases ; complications ; pathology ; Bone Diseases, Infectious ; complications ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; complications ; microbiology ; Male ; Middle Aged ; Necrosis ; Osteomyelitis ; complications

Age-related macular degeneration ( AMD ) is a kind of age-related blinding degenerative fundus lesions, totally about 30 million patients suffering from AMD all over the world, with about 500 000 people blind for it yearly. As the development of economy and the aging of the population intensified, incidence of AMD indicates a trend of rising year by year, being the third major cause of blindness in our country. At present, the pathogenesis of AMD is not fully clear, as reported it may be related to oxidative stress, inflammatory immune response, VEGF and genetic manipulation. Clinical treatments mainly include photodynamic therapy, drug therapy, radiation therapy, laser photocoagulaory operation, the pupil warm treatments, Chinese medicine and intravitreous injection VEGF antagonists such as Ranibizumab, Conbercept and so on. ln this issue, we mainly expound on the progress in the epidemiological studies of AMD, especially elaborate the progress made on genetic manipulation in recent years.

The microbial populations and community structure characterization technologies of the enhanced biological phosphate removal system were reviewed comprehensively in this paper, and their future research directions were outlined.

The kinase domain of receptor tyrosine kinase(RTK) ErbB2 was expressed fused with GFP in Pichia pastoris. Recombinant expression vector pPIC3.5K was constructed in Escherichia coli TOP10. The right P. pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of recombinant vector, and then induced by methanol in baffled shake bottles. The strain with highest protein yield was scaled up in a 5 L fermentor. Recombinant protein was analyzed with tyrosine kinase assay after Ni2+ affinity chromatograph. Results showed that the 100 kD recombinant protein with tyrosine kinase activity was successfully expressed in P. pastoris.

Background Researches showed that wild-type p53(wtp53)and Rb94 genes inhibit the growth of retinoblastoma(RB),and these genes are involved in signal pathway in the induction and maintenance of cellular senescence.Thus the combination of two genes to inhibit growth of RB is concerned.Objective This study was to observe the inhibitory effect of the co-transfection of Rb94 and wtp53 gcnc into subretina on RB with ultrasound microbubble.Methods HXO-Rb44 suspension was subretinally injected to establish the RB model in 40 SPF female Balb/c nude mice.The RB models were randomized into model control group,wtp53 group,Rb94 group and wtp53+Rb94 group,and 0.1 ml relevant gene microvesicle suspension was injected via caudal vein in the different groups,but no any gene was used in the model control group.Seven days after modeling,followed by 0.5 W/cm2ultrasonic wave irradiated the eyeballs immediately for 4 seconds ×2 times and interrupted for 24 seconds.Eyeballs were extracted 7 days after gene transfection,and the expressions of wtp53 mRNA and Rb94 mRNA in tumor tissuc were detected by RT-PCR,and wtp53 and Rb94 protein in tumor tissue were assayed using Western blot.Immunochemistry was used to exam the VEGF expression,and TUNEL was used to evaluate the apoptosis of the tumor cells.Results The model successful rate after HXO-Rb44 suspension was 80％ (32/40)and obvious malformation cells were seen under the light microscope.In 7 days after gene transfection,no response band for wtp53 mRNA and Rb94 mRNA were found.The relative expression valuc of wtp53 mRNA was 0.65±0.07 in the wtp53 group,and that in wtp53+Rb94 group was 0.32±0.02,showing a significant difference between them (t =11.743,P =0.000).Rb94mRNA relative value was 0.42 ±0.03 in Rb94 group,and that in the wtp53 + Rb94 group was 0.23 ± 0.03,with a significant difference(t=5.041,P=0.001).The response bands of wtp53 and Rb94 proteins were seen in wtp53group,Rb94 group and wtp53+Rb94 group.Immunochemistry showed that the positive reactive intensity for VEGF in tumor tissue was obviously weaker in wtp53+Rb94 group than that in the wtp53 group,Rb94 group and model control group.Apoptotic index(Al) was 37.35±2.14 in the wtp53+Rb94 group,showing a significant increase in comparison with model control group (0.46 ± 0.05),wtp53 group (5.05 ± 0.80) and Rb94 group (6.43 ± 1.02) (t =-34.395,-28.206,-26.006,P＜0.01).Conclusions Ultrasound microvesicle enable double gene transfecting into RB tumor tissue,and Rb94 gene cooperation with wtp53 gene enhance the inhibitory effect on RB by promoting the apoptosis of RB cells.

Objective To approach the evaluative effect of respiratory variation of superior vena cava peak flow velocity measured using transthoracic echocardiography (TTE) on fluid responsiveness in patients with mechanical ventilation.Methods A prospective cohort study was conducted.All mechanical ventilated critically ill patients whose fluid therapy was planned due to hypovolemia in Department of Critical Care Medicine of Beijing Tongren Hospital of Capital Medical University from April 2011 to April 2013 were enrolled.Volume expansion was performed with 500 mL Linger solution within 30 minutes.Patients were classified as responders if pulse pressure variation (PPV) increased ≥ 13％ before volume expansion.The respiratory variation in superior vena cava peak velocity was calculated as the difference between maximum and minimum values of velocity in peak A,peak S and peak D over a single respiratory circle,and their variations (ΔA,ΔS,ΔD) were also calculated.The receiver operating characteristic curve (ROC curve) was plotted to assess the evaluative effect of respiratory variation of superior vena cava peak velocity on fluid responsiveness.Results Twenty-seven patients were enrolled in this study.Volume expansion increased PPV ≥ 13％ happened in 14 patients (responders).The velocity of superior vena cava in peak A,peak S,peak D was significantly increased after volume expansion compared with that before volume expansion in responders [peak A (cm/s):34.6 ± 2.2 vs.31.3 ±2.1,t=-2.493,P=0.027; peak S (cm/s):39.1 ± 1.3 vs.35.3 ±2.1,t=-2.564,P=0.024; peak D (cm/s):28.1 ± 1.2 vs.23.3 ± 1.4,t=-4.995,P=0.000],but there was no significant difference in ΔA,ΔS and ΔD between before and after volume expansion.The ΔA,ΔS and ΔD were positively correlated with PPV (r＝0.040,P=0.854; r=0.350,P=0.074; r=0.749,P=0.000).The area under ROC curve (AUC) of peak S was 0.36 [95％ confidence interval (95％CI):0.11-0.52],but the AUC of ΔS was 0.68 (95％CI 0.47-0.89),the AUC of peak D was 0.41 (95％CI 0.19-0.63),but the AUC of ΔD was 0.95 (95％CI 0.86-1.00),so the aberration rate of superior vena cava in respiration was better than the flow rate in superior vena cava.When the cut-off value of ΔS was 20.7％ for predicting fluid responsiveness,the sensitivity was 78.6％ and the specificity was 61.5％.When the cut-off value of ΔD was 12.7％ for predicting fluid responsiveness,the sensitivity was 92.0％ and the specificity was 92.3％.Conclusion Respiratory variations in superior vena cava peak velocity measured by TTE could assess fluid responsiveness in patients with mechanical ventilation.

Objective To observe clinical efficacy of treating acute facial neuritis with routine acupuncture treatment and tube moxibustion therapy. Methods Totally 100 cases were divided into observation group (50 cases) and control group (50 cases) with random number table method. Both groups were treated with acupuncture on accupoints of Yifeng (SJ17), Dicang (ST4), Jiache (ST6), Xiaguan (ST7), Yingxiang (LI20), Taiyang (EX-HN5), Cuanzhu (BL2) of affected side, Hegu (LI4) of unaffected side. On the basis of routine treatment, observation group was given electronic tube moxibustion therapy, inserting the ipsilateral external auditory canal about 1 cm and waiting for burning out, 20 min each time. Control group received TDP to irradiate ipsilateral ear, 20 min each time. Treatment for both groups were once a day, 5 times as a treatment course, 2 d between each course, for 4 courses. House-Brakmann facial nerve functional index, temperature differences on face and retroauricular skin and ear pain duration were observed. Clinical efficacy was evaluated. Results One and two cases were lost in observation group and control group, respectively. The total effective rate of clinical efficacy was 91.67% (44/48) in the observation group and 75.51% (37/49) in the control group, with statistical significance (P<0.05). Compared with before treatment, House-Brakmann facial nerve functional index in both groups improved (P<0.05), and the observation group was better than the control group (P<0.05). Compared with before treatment, temperature differences on face and retroauricular skin in both groups decreased, and the observation group was better than the control group (P<0.01). The ear pain duration in the observation group was shorter than the control group (P<0.05). Conclusion Tube moxibustion therapy combined with routine acupuncture can improve the symptoms of acute facial neuritis and promote clinical efficacy.

Objective To evaluate the effects of 131 adrenoceptor anfisense on blood pressure and?1 adrenoceptor mRNA and protein levels in 2 kidney 1 clip(2K1C)rats.Method 2KIC hypertensive rots were produced by clipping renal artery of SD rats.Liposome/AS-ODNs 2.0 were tested intravenously in rats with 2KIC hypertension.Animals were divided into 5 groups(n=18 in each group):?1-AS-ODN group,?1-IN-ODN group,2K1C group,Sham group and SD group.Blood pressure was measured by tail-cuff method,the levels of myocardial?adreneceptor mRNA and protein were tested by RT-PCR and binding assay.Results On the basis of the magnitude and duration of hypotension,?1-AS-ODN decreased blood pressure by 39 mmHg at the most for 4 weeks.Compared with the 2KIC group,?1-AS-ODN did not significantly change the levels of myocardial?1 adrenoceptor mRNA but significantly decreased the levels of myocardial?1 adrenoceptor protein at 2,7,30 days (P

OBJECTIVE: To study the clinical value of polysomnography (PSG) and ApneaGraph (AG200) in the diagnosis evaluation of obstructive sleep apnea hypopnea syndrome (OSAHS). METHOD: From January to December in 2012, 67 OSAHS patients diagnosed by PSG were examined by AG200. The apnea hypopnea index (AHI), hypopnea index (HI), apnea index (AI), the lowest oxygen saturation (LSaO2) was were detected and the results were analyzed statistically. RESULT: Significant differences were observed in AI, AHI, LSaO2 between AG200 and PSG (P < 0.05, respectively). No statistically significant difference was found in HI. The differences in HI between AG200 and PSG were not significant (P > 0.05). AHI, HI and LSaO2 was were significantly correlated between AG200 and PSG (r = 0.870, 0.743, 0.374, 0.716, P < 0.01). CONCLUSION AG200 could not replace PSG but could identify the level of upper airway obstruction.
Female ; Humans ; Male ; Polysomnography ; methods ; Sleep Apnea, Obstructive ; diagnosis ; physiopathology