Objective To observe the interferon-γ(IFN-γ),interhukin-2(IL-2)levels of Th1 cytokine and IL-4、IL-10 levels of Th2 cytokine in serum and culture supernatants of splenic cells of the rats in the prevention of experimentaI autoimmune uveoretinitis(EAU)by oral tolerance. Methods 72 Lewis rats were randomly divided into EAU group,oral tolerance group(which including 10 μg、100 μg、1 mg、10 mg of S antigen group respectively)and control group,12 rats in each group.The animal model of EAU was induced by immunization with S antigen(50 μg)and Freund's complete adjuvant.Oral tolerance 10 μg、100μg、1 mg and 10 mg group were fed with 1 ml mixture of 10 μg、100μg、l mg、10 mg S antigen and 1 mg trypsin inhibitor respectively by intuhation,once the other day,totally 7 times,and then induced EAU according to above methods;control group was fed with 1 ml mixture of phosphate buffered saline and 1 mg trypsin inhibitor,once the other day,totally 7 times,and then induced EAU.The clinical manifestation of EAU in the eye were recorded,the eyeballs were enucleated at the peak of EAU,followed by pathological grading.Meanwhile the serum was colleced;splentic cells were separated and cultured to collect the supernatant.Cytokine levels of IFN-γ,IL-2,IL-4 and IL-10 in serum.cultured supernatant of splenic cells were determined by enzyme-linked immunosorbent assay(ELISA). Results Compared with EAU and control group,the levels of IFN-γ and IL-2(Th1 eytokine)in the serum in 100 μg and 1 mg group were decreased while the levels of IL-4 and IL-10(Th2 eytokine)were inereased,the diffefences were statistically significant(F=51.9,68.8,35.7,7.5,P＜0.01).Compared the levels of Th1 and Th2 cytokines in the serum in 10 μg,10 mg group with EAU and control group.the differences were not statistically significant.In 100 gg、1 mg group,the levels of IFN-γ and IL-2(Th1 cytokine)in the culture supernatant of splenic cells were decreased while the Ievels of IL-4 and IL-10(Th2 cytokine)were increased,compared with EAU and control group,the differences were statistically significant(F=57.1,15.6,33.1,167.7,P＜0.01).Compared the levels of Th1 and Th2 eytokine in the culture supernatant of splenic cells in 10 μg、10 mg groups with EAU and control group,the difference are not statistically significant. Conclusions In the process to prevent EAU by oral intake,the levels of IFN-γ and IL-2 (Th1 cytokine)were decrease while the levels of IL-4 and IL-10(Th2 cytokine).Oral administration with tOO high or low dose of the antigen can not prevent EAU as well as the cytokine levels do not change obviously.Cytokines has played an important role in the prevention of EAU.

Objective To observe the efficacy of the anti-tumor necrosis factor-α monoclonal antibody(TNF-α MCAb)in the treatment of experimental autoimmune uveoretinitis(EAU). Methods EAU animal models were induced by interphotoreceptor retinoid-binding protein(IRBP)R16 peptide with immunization.The rats were divided into 2 groups according to the injection times.TNF-α MCAh was administered intravenously on day 6 or 4,6 and 8 post-immunization respectively,and then to observe the clinical expression by slit-lamp microscope.Meanwhile,take the rats which did not accept TNF-α MCAb as conttol group.Delayed type hypersensitivity(DTH)responses were measured on day 13 postimmunization of IRBP R16;the rats were killed on day 14 post-immunization of IRBP R16,and then enucleated the eyes for histopathological examination.To detect the cytokine level of IFN-γ,IL-4 in serum and IFN-γ in aqueous humor by enzyme-liked immunosorbent assay(ELISA)on day 14 postinjection.The hyperplasia responses of antigen specific lymphocyte of draining lymph node cells were detected. Results The TNF-αMCAb group had mitigated ocular inflammation and decreased pathological grades compared with the control group;the IFN-γ concentrations in aqueous humor and serum were decreased,IL-4 was increased in serum;DTH responses were decreased;the hyperplasia responses of draining lymphocytes to IRBP R16 peptide were decreased,all the differences were statistically significant(P＜0.01).The rats accepted TNF-α MCAb thrice had much better curative effect than the rats injected once(P＜0.05). Conclusions Injection of TNF-α MCAb can inhibit ocular inflammation and specific immune cells of EAU remarkably and change the Th1/Th2 balance.Many times injections of TNF-α MCAb were more effective than once.

BACKGROUND:Artificial cervical disc prosthesis simulates range of motion and buffer shock function of normal intervertebral discs. Clinical experiments verify that artificial cervical disc prosthesis material has good biocompatibility and mechanical characteristics. OBJECTIVE:To evaluate artificial cervical disc replacement and zero-profile interbody fixation and fusion system for multilevel cervical disease in 2-year folow-up. METHODS:Artificial cervical disc replacement and zero-profile interbody fixation and fusion system were used to treat 42 patients with multilevel cervical disease. The patient presented typical symptoms and signs of spinal cord or nerve root compression. There were 18 cases of cervical myelopathy, 15 cases of nerve root cervical spondylosis and 10 cases of mixed type of cervical spondylosis. After treatment, mean operation time, blood loss and reoperation rate were measured. Postoperative complications, disability index of neck function, visual analog scale, function unit range of corresponding surgery segments of the cervical spine, Cobb angle of C2-C7 vertebral body, range of motion of adjacent segment of proximal and distal vertebral bodies were observed and clinical outcomes were evaluated. RESULTS AND CONCLUSION: Al cases finished the operation and were scored at various time points. After treatment, radiating pain of shoulder and neck and upper extremity were remarkably lessened. Numbness and sensory loss symptoms disappeared obviously. Quality of life elevated noticeably. Visual analog scale and the disability index of neck function score were decreased in final folow-up compared with pre-treatment (P < 0.001). C2-C7 vertebrae Cobb angle, FSU angle, range of motion of proximal surgery adjacent segment and range of motion of the distal surgery adjacent segment were elevated compared with pre-treatment (P < 0.001). These data indicate that cervical spondylosis was improved after treatment. Each index of cervical spondylosis after artificial cervical disc replacement and zero-profile interbody fixation and fusion system was reconstructed to different degrees.

Objective To determine the distribution of IL-4, IL-17, IFN-γ in the lacrimal gland lesions and submaxillary lymph nodes of MRL/Ipr mice-a model for idiopathic dacryoadenitis, and investi-gate the possible role of T-belper cells in the disease. Methods Frozen sections of lacrimal glands and sub-maxillary lymph nodes from MRL/lpr and BALB/c mice age 3 months were stained with antibodies to the cy-tokines IL-4, IL-17 and IFN-γ, then calculated the numbers or proportions of positive cells respectively,double staining for CD4 and each cytokine was taken in lymph nodes of MRL/Ipr mice additionally. Results The major cytokine in lacrimal gland lesions is IL-4, which contrasts to little IL-17 or IFN-γ/stained(P <0.05), the numbers of cells staining for each cytokine in submaxillary lymph nodes elevate in MRL/lpr mice, and the expressions of IL-4 and IL-17 are greater than IFN-γ significantly. Conclusion Th2 may play an important role in the development of local dacryoadenitis and systemic abnormality in MRL/lpr mice,the role of IL-17 maybe different in lacrimal gland lesions and lymph nodes which need more research.

ObjectiveTo investigate the effects of astragalin on pulmonary edema after acute cervical spinal cord injury in rats.MethodsA total of 200 adult Wistar rats weighing 240-250 g were randomly divided into five groups:astragalin group,low concentration astragalin group,physiological saline group,blank group and sham group,with 40 rats in each group.The rats with cervical spinal cord injury were induced at C7 by modified Allen' s method,with the dropping weight of 10 × 2.5 g · cm.In the sham group,the laminas were removed only,leaving spinal cord at the C7 intact.Each group was further divided into four time points:24 hours,3 days,1 week and 2 weeks after the modeling,with 10 rats in each time point,according to the specimen collection time.Rats were sacrificed at different time points to observe the pathological change of the lung tissue using optical microscope,measure the lung wet/dry weight ratio (W/D) and protein concentrations of the serum and bronchoalveolar lavage fluid and calculate the lung W/D and lung permeability index (LPI).ResultsAt the same instant,the W/D and LPI in the astragalin group and low concentration astragalin group were lower than those in physiological saline group and blank group,with the lowest value in the astragalin group at day 3 after injury ( P ＜0.05 ).ConclusionsRats with acute cervical spinal cord injury may cause pulmonary edema,which can be efficiently alleviated through early use of astragalin.

Objective To investigate the expression of lung keratinocyte growth factor receptor (KGFR) in rats with acute spinal cord injury (ASCI) in different time points and its role in lung edema.Methods Thirty-two adult Wistar rats weighing 240 g to 260 g were assigned to experimental group (n =16) and control group (n =16) according to the random number table.Each group consisted of time points of 24 hours,3 days,1 week and 2 weeks after the modeling (4 rats per time point).A rat model of ASCI in experimental group was induced at C7 segment by dropping a weight of 10 g from the height of 2.5 cm (Allen' s method).In control group,laminas were removed only,leaving spinal cord at C7 intact.Rats were sacrificed at each time point for measurement of lung wet/dry weight ratio,Western blot analysis of expression of lung KGFR protein and RT-qPCR detection of lung KGFR mRNA expression.Results After ASCI in rats,the expressions of lung KGFR protein and mRNA began to drop at 24 hours (0.23 ±0.06,0,012 1 ±0.002 3),reached the trough at 3 days (0.17 ±0.04,0.008 5 ±0.001 7)and picked up at 1 week.Expression of lung KGFR mRNA in experiment group showed statistically significant difference from that in control group at 24 hours and 3 days (P ＜ 0.05),whereas in each time point the difference of KGFR protein expression between experiment and control groups was statistically significant(P ＜0.05).Variation trend of KGFR expression was in parallel with the severity degree of pulmonary edema.Conclusion Lung KGFR presents significant down-regulation in ASCI rats and this may be associated with the development of pulmonary edema after ASCI.

Objective To determine the effectiveness of systolic blood pressure gradient between the right arm and the right leg(SBPG) tests in the diagnosis of neonatal aorta anomaly(AoA).Methods The SBPG of infants suspected of having critical congenital heart diseases were collected prospectively,who were admitted to Guangdong General Hospital from January 2013 to December 2015.The results of SBPG test were compared with those of echocardiography or cardiac computed tomography (golden standard).The rates of true positive,true negative,false positive,false negative were calculated under cutoff values of 5 mmHg(1 mmHg =0.133 kPa),10 mmHg,15 mmHg and 20 mmHg,respectively.Receiver operating characteristic (ROC) curves was used to compare tests of different cutoff and the areas under the ROC curve were also calculated.Results Among 664 enrolled infants,67 cases were confirmed by golden standard test.The systolic blood pressure in the right arm,the legs and SBPG in AoA group and non-AoA group were (88.0±20.4) mmHgvs.(73.4±9.3) mmHg (P＜0.01),(66.1 ±10.1) mmHg vs.(69.0 ±9.7) mmHg(P＞0.05) and (22.6±17.8) mmHgvs.(2.3 ±4.8) mmHg(P ＜0.01),respectively.In these patients,31 cases(46.3％),31 cases(46.3％),27 cases(40.3％) and 21 cases(31.3％) were diagnosed of AoA,and 36 cases(53.7％),36 cases (53.7％),40 cases(59.7％),and 46 cases (68.7％) were missed by SBPG tests of 5 mmHg,10 mmHg,15 mmHg and 20 mmHg,respectively (P ＜ 0.01).The rates of true negative among those groups were 94.1％,99.5％,99.7％ and 100.0％,and the areas under ROC curve were 0.656,0.722,0.695 and 0.657,respectively (P ＜ 0.01).Conclusions Almost half of AoA infants could be screened out by SBPG test.The cutoff of 10 mmHg could probably be used to screen potential AoA infants,with higher true positive rate and lower false positive rate.

AIM: To study the inhibitory effects of nm23-H1 gene on proliferation and invasion of human lung adenocarcinoma A549 cell line. METHODS: Recombinant eukaryotic expression vector pcDNA3.1-nm23-H1 containing full length of human nm23-H1 cDNA was constructed and transfected into a human lung adenocarcinoma A549 cell line by lipofectamine. Cell strain that expressed nm23-H1 stably was screened out by G418 and named pcDNA-nm23-A549. Expression of nm23-H1 was identified by RT-PCR and immunohistochemistry. Growth curves were drawn to detect the inhibitory effects on cell proliferation. Cell cycle of pcDNA-nm23-A549 was examined by flow cytometry. Atomic force microscopy was used to observe the filopodia on the surface of the cells. RESULTS: Introduction of nm23-H1 obviously inhibited the proliferation of A549. Expression of nm23-H1 did not induce apotosis in A549 cells but increased the percentage of phase G_1 cells and decreased phase S cells. Meanwhile, phase G_1 to phase S transition was restrained. Filopodia in the cell surface was much fewer and its structure changed in cells transformed. CONCLUSION: nm23-H1 is capable of inhibiting A549 proliferation and decreasing its metastatic ability, probably by interfering with cell cycle and cell surface structure.

Objective To investigate the therapeutic effects of thymopentin(TP-5)for the experimentsl autoimmune encephalomyelitis(EAE)in rats.Metbods The EAE model wss established in wistar rats immunized with fresh guinea pig spinal cord homogenate(GPSCH)and complete Freund's adjuvant (CFA).Wistar rats were divided randomly into five groups:normal control group,EAE group,dexamethaSone(DXM)group,low dose TP-5 treated group,hish dose TP-5 treated group.The levels of IL-12 and IL-10 in serum of wistar rats were detected by sandwich-ELISA on day 7,14 and 21 post immunization.Resuits Morbidity and clinical score of low dose TP-5 treated group and DXM group were significantly lower than those of EAE group and hish dose TP-5 treated group(P＜0.01).Morbidity and clinical score of DXM group were significantly lower than those of low dose of TP-5 treated group(P＜0.01).The levels of IL-12 of EAE group,DXM group,low dose TP-5 treated group and high dose TP-5 treated group were significantly higher than of normal control group at each time point(P＜0.01).The levels of IL-12 of DXM group and low dose TP-5 treated group were significantly lower than that of EAE group(P＜0.01),meanwhile the levels of IL-10 of DXM group and low dose TP-5 treated group were significantly higher than that of the rest groups on day 14 and 21 post immunization(P＜0.01).Conclusion TP-5 has protective effects on EAE,and its functional mechanism may be associated with down.regulation of IL-12 as well as up-regulation of IL-10,so as to reverse the imbalance of TH1/TH2 responses by bidirectional regulation.

OBJECTIVETo investigate the expression pattern of the cystatin-related epididymal spermatogenic (Cres) gene in mouse testes and epididymis during postnatal development.

METHODSSemi-quantitative RT-PCR was used to detect the Cres mRNA level in postnatal mouse testes and epididymis on day 14, 20, 22, 28, 35, 49, 70 and 400.

RESULTSLow-level Cres mRNA was detected on day 14 in both the testes and the epididymis. The expression of the Cres gene increased gradually with the development of the mouse, and it reached the peak on day 70 in the testes and on day 400 in the epididymis.

CONCLUSIONThe time-dependent expression pattern of the Cres gene in postnatal mouse testes and epididymis suggested that the Cres gene might be involved in the regulation of spermatogenesis and sperm maturation.

Age Factors ; Animals ; Blotting, Northern ; Cystatins ; genetics ; Epididymis ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; analysis ; Testis ; metabolism