Background and purpose: Urokinase-type plasminogen activator receptor is related to invasion and metastasis of tumor. Inhibition of uPAR expression in tumor cells results in reducing its metastasis. This study was aimed to construct an expression vector with short hairpin RNA (shRNA) of uPAR, which could pave the way for RNAi-mediated tongue squamous cell carcinoma therapy. Methods: Genome sequences of uPAR gene were retrieved from Genhank and cDNA was designed to code expression of shRNA for uPAR gene. The cDNA was synthesized and inserted into the eukaryotic expression vector pWH1, and the recombinant pWH1-uPAR expression vector was identified by enzyme cutting method. Then, pWH1-uPAR expression vector was transfected into tongue squamous cell carcinoma Ts cells by Lipofectomine 2000. At last, the expression of uPAR in Ts cells transfected with pWH1-uPAR expression vector was observed by RT-PCR, immunocytochemistry staining and Western blot. MTT assay was performed to measure the proliferation of Ts cell. Results: The uPAR shRNA eukaryotic expression vector was successfully constructed. Compared with Ts cells and Ts cells transfected with plasmid pWH1, the Ts cells transfected with pWHI-uPAR expression vector showed a lower mRNA and protein expression of uPAR. The inhibition rate of proliferation was 32.9% of Ts cells by transfected with pWHl- uPAR. Conclusion: The constructed uPAR shR.NA expression vector could inhibit the expression of uPAR in tongue squamous cell carcinoma, which may be helpful for further research on the function of uPAR and provide effective methods for therapy of tongue squamous cell carcinoma.

Objective Curcumin and MS-275,an inhibitor of histone deacetylase (HDAC),are promising anti-tumor agents. The aim of present study was to investigate the mechanisms of apoptosis induced by combined use of MS-275 in low dosage and curcumin in prostate cancer cell line DU145. Methods MTT assay was used to evaluate the lethal effect on DU145 cells by solitary use of MS-275 and or combined with curcumin. The changes in cell life cycle was detected by flow cytometry. The expressions of the survival signaling pathways were determined by Western blotting. Results Solitary application of MS-275 or curcumin may inhibit the growth of DU145 cells in a time and dose-dependent manner. The combination of MS-275 (1?mol/L) and curcumin (20?mol/L) exhibited obvious cytotoxic effect on cell viability,which was only 45.9% 48h after the combined treatment. Cell cycle assay showed that the combination of MS-275 and curcumin resulted in obvious appearance of sub-G1 phase in DU145 cells,implying that the cell apoptosis had been induced. The results of Western blotting showed that after treatment of MS-275 or curcumin singly,the phosphorylation level of Akt and ERK kinase declined slightly,however,when MS-275 and curcumin were used together,there appeared a prominent inhibitory effect on Akt and ERK kinase,indicated by a sharp decline of their phosphorylation level,and at the same time,the level of cleaved PARP,a hallmark of apoptosis,was increased in DU145 cells. Conclusion Combined use of MS-275 and curcumin may exert a synergistic cytotoxic effect on the viability of DU145 cells,and exhibit an inhibitory activity on Akt and ERK kinases to induce apoptosis.

The cerebral blood flow of infant is effected by physiological and pathological factors.As the cerebrovascular autoregulation of neonates is poor,in pathological cases,especially when hypoxemia and hypercapnia impaired regulation of its own,lead to changes in cerebral blood flow,then resulting in severe brain injury.It has made enormous progress in the research on the changes of cerebral blood flow in newborns in recent years.In normal infants,cerebral blood flow velocities is positively correlated to gestational age and body weight,and increase gradually with day-age in the first week after birth.The cerebral blood flow of newborn with brain injury can present as insufficiency,over-perfusion or low speed high-resistance earlier and high speed low-resistance later.Different results may be related to the duration and severity of asphyxia,but all of those are signs of damage of self-regulatory function of cerebral blood flow.Cerebral hemodynamic change is the important pathogenesis mechanism of brain injury.

Background The rejection following keratoplasty still is a leading cause of corneal transplantation failure.Studies showed that the interleukin-22 (IL-22) ,one of the effector molecules of T helper cell 17 (Th17) participated on the rejection after heart,liver and bone marrow transplantation.However,the effect of IL-22 on corneal graft rejection is not well understood.Objective This study was to investigate the expression of IL-22 mRNA in the corneal grafts and the role of IL-22 in the immune rejection after corneal transplantation in rats.Methods Seventy-two Wistar rats were randomized into autologous keratoplasty group,allograft keratoplasty group and anti-rejection group,and other 4 normal Wistar rats served as normal control group.Autologous keratoplasty was operated on the Wistar rats of the autologous keratoplasty group,and allograft keratoplasty were carried out with the 24 SD rats as donors and 48 Wistar rats as recipients.Tobramycin and dexamethasone eye drops were topically administrated after autologous keratoplasty for 2 weeks in the anti-rejection group.The experimental eyes were examined by slit lamp microscope after surgery and graft survival was evaluated based on the rejection scoring criteria of Larkin.Intergroup accumulated survival rates of grafts were compared using Kaplan-Meier analysis.Histopathological examination of grafts was carried out in 5 and 14 days after operation respectively,and the related expression levels of IL-22 mRNA and aryl hydrocar-bon receptor (AhR) mRNA were carried out by real-time fluorescence quantitative PCR.The feeding and use of the experimental animals followed the Guangdong provincial regulations on the management of experimental animals.The experimental design was approved by the ethics committee of Southern Medical University.Results The median survival time of grafts in the allograft keratoplasty group was 10 days,and that in the anti-rejection group was 17 days,showing a significant survival extention in the anti-rejection group (x2=16.442,P =0.000).Significant differences were found among the 4 groups in the related expression levels of IL-22 mRNA in both 5 days and 14 days after surgery (postoperative 5 days : F=2.44,P =0.00;postoperative 14 days: F=267.92, P =0.00), and the related expression levels of IL-22 mRNA were remarkably higher in the allograft keratoplasty group than those in the anti-rejection group at different time points (postoperative 5 days :9.70±0.35 vs.0.46±0.21;postoperative 14 days : 23.12 ± 1.89 vs.3.14±0.94) (both at P＜0.05).The related expression levels of AhR mRNA in the grafts were considerably different among the 4 groups (postoperative 5 days : F =395.73, P =0.00;postoperative 14 days : F =942.37, P =0.00) , and the expression levels were significantly elevated in the allograft keratoplasty group compared with the anti-rejection group at various time points (postoperative 5 days:2.52±0.32 vs.1.89±0.10;postoperative 14 days:7.20±0.25 vs.2.60±0.17) (both at P＜0.05).Conclusions The expression level of IL-22 RNA up-regulates in the grafts with immuno-rejection.Topical administration of tobramycin and dexamethasone eye drops inhibits the rejection after keratoplasty.AhR plays a regulative role to the expression of IL-22 in rats after keratoplasty.

0.05).Conclusion DTI can noninvasive detect the potential disorder of corpus eallosum in vivo,thus providing useful information to differentiate the cerebral ischemia disease from multiple sclerosis.

Objective To clarify the role of B7-H1 in the immune privilege after corneal transplantation in homogeneity variant mice.Methods We established the experimental animal model of allograft mice by using C57BL/6 mouse as donor and Balb/c mouse as recipient.We allocaated the mice with long time survival (＞50 days) corneal graft into survival group,mice with rejection occurring in 50 days into rejection group,and normal C57BL/6 mice into control group.The transplanted corneal grafts were obtained for future reference at the 8th week after transplantation in survival group,and the time of rejection in rejection group.The expression of B7-H1 mRNA was detected by using immunohistochemistry and real time quantitative PCR (RT-PCR),and the relationship between B7-H1 and the immune privilege after corneal transplantation was analyzed. Results The B7 H1 mRNA was highly expressed in epithelium and endothelium of corneal grafts both in survival and control group,in comparison to an obviously lower expression in rejection group.The relative expression level of B7-H1 mRNA was 200.0 ± 11.5 in survival group,44.7 ± 10.8 in control group,and 6.9 ± 12.0 in rejection group,respectively. There were statistically significant differences among the three groups (F=241.164,P＜0.01 ).The was a significant correlation between the level of B7-H1 mRNA and occurrence rate of rejection in corneal graft (P＜0.01 ).Conclusion The results suggest that the immune privilege after corneal transplantation might be mediated by B7-H1,which plays an important role in maintaining the state of corneal immune privilege.

Objective To understand the iodine deficiency disorders in Xunhua county Qinghai province, and to provide reference values for setting up prevention and control strategy. Methods According to historical data of Xunhua county, 1 township center elementary school and 1 village elementary school were sampled in Qingshui, Baizhuang, Hongqi, Jiezi and Galeng townships. A total of 9 elementary schools were investigated (Caleng has only 1 elementary school). Fourty children aged 8-10 years were checked thyroid size by palpation in every school. Twenty children were chosen to assay their urinary iodine and edible salt iodine in every school. Thirty 5-grade students were surveyed by questionnaire on health education in every school. Ten housewives were inquired prevention knowledge on iodine deficiency disorders and salt iodine level was tested in every village. Results Goiter rate was 7.9%(30/378) in children aged 8-10 years, with Baizhuang and Galeng the highest (11.9%,5/42), and Qingshui the lowest (2.4%, 1/42). The median was 196.5 μg/L in children aged 8-10 years, and urinary iodine level lower than 50 μg/L was accounted for 5.3% (11/208), but that value was 25.0% (5/20)in Galeng. The average qualified rate of using edible salt was 91.4%(159/174), and that rates were 79.0%(15/19), 83.3%(15/18) and 89.5%(17/19) in Chajia, Tuanjie and Minzhu villages, respectively. The average score were 2.8, 2.1 in health education survey of the 5-grade students and housewives. Conclusions After years effort in prevention and control of iodine deficiency disorders, remarkable progress has been made in Xunhua County. The monitoring of iodized salt and the knowledge spreading are still need to be strengthened.

ObjectiveTo explore the role of ssomatosensory evoked potential (SEP) in evaluating and predicting the lower limb motor function of patients with acute cerebral infarction.Methods19 patients (13 male, 6 female) with acute cerebral infarction diagnosed according to the standard of Chinese Classifications of the Cerebral Vascular Disease were chosen as the case group and 18 patients (11 male, 7 female) with non-nerve system injury were taken as the control group. All patients of two groups were received SEP test within 7 days after onset and latency and amplitude values of SEP of two groups were compared. To the case group, SEP was tested again at 30th day after admission, the Fugl-Meyer assessment (FMA) was also carried out at same times of two SEP tests, and the correlation of SEP and FMA scores was analyzed.ResultsThe difference of latency and amplitude value of SEP test between the case group and control group was significantly (P<0.01). The latency value of SEP in lower limb was correlated with FMA scores (P<0.01). But the amplitude value was no difference between the onset and 30th day after admission in the case group.ConclusionSEP is sensitive in the acute stage of stroke. SEP has significant value for predicting the prognosis of patients with acute cerebral infarction and correlates with the clinical FMA scores. The value of amplitude is not sensitive.

Adult ; Aged ; Antigens, CD20 ; metabolism ; Diagnosis, Differential ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Humans ; Immunohistochemistry ; Leukocyte Common Antigens ; metabolism ; Lymphoma, B-Cell ; genetics ; metabolism ; pathology ; surgery ; Lymphoma, Extranodal NK-T-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; surgery ; Male ; Middle Aged ; Tonsillar Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Tonsillectomy ; Tonsillitis ; pathology ; Young Adult

OBJECTIVETo investigate the changes in the levels of monocarboxylate transporter-2 in spinal cord horn in a rat model of chronic inflammatory pain.

METHODSMale SD rats weighting 180 - 220 g were randomly divided into two groups(n = 48): normal saline group (NS group), complete Freund's adjuvant group (CFA group). Rats were given injections of CFA 100 µl in left hind paw in group CFA, and an equal volume of saline was given injection in group NS. Mechanical withdraw threshold(MWT) and thermal withdraw latency(TWL) were measured at before injection(T0 and 3 h, 1 d, 3 d, 7 d, 14 d, and 21 d after injection(T1-7). Four rats were chosen from each group at T0-7 and sacrificed, and L4-5 segments of the spinal cord horn were removed for measurement of the expression of monocarboxylate transporter-2 by Western blot analysis.

RESULTSIn CFA group, mechanical hyperalgesia and allodynia appeared on the 3 h after CFA injection, then until the day 14. The expression of monocarboxylate transporter-2 in the spinal dorsal horn of rats in CFA group was significantly higher than that in normal control group at T1-6(P <0.05). The protein level of monocarboxylate transporter-2 was apparently correlated with MWT and TWL(P <0.01 and P <0.05) in CFA group.

CONCLUSIONThe level of monocarboxylate transporter-2 in spinal dorsal horn is significantly increased in a rat model of chronic inflammatory pain and the change may involve in the formation and maintenance of central sensitization in spinal cord of chronic inflammatory uain.

Animals ; Disease Models, Animal ; Freund's Adjuvant ; Hyperalgesia ; chemically induced ; Inflammation ; chemically induced ; metabolism ; Male ; Monocarboxylic Acid Transporters ; metabolism ; Pain ; chemically induced ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; physiopathology