Objective: To investigate the pharmacokinetics of rhein (RH) in rat plasma and kidney simultaneously. Methods: An HPLC method was developed to determine the concentration of RH in rat plasma, which was corrected with protein binding. Renal microdialysis (MD) technique was employed to collect dialysate sample continuously. Then the MD samples were detected by HPLC-MS/MS method. Results: RH was distributed into kidney of rats rapidly after administration. The maximum concentration of RH in rat kidney was reached at 1 h followed by continuously decreased. During the whole process, the concentration of RH in rat kidney was significantly lower than that in plasma after administration. The MRT of RH in plasma and kidney was (87.217 ± 29.889) and (122.387 ± 12.521) min, respectively. The areas under the curve (AUC) of RH in plasma and kidney were (114.236 ± 45.585) and (16.596 ± 1.732) μg·min/mL, respectively. The kidney penetration was 0.161 ± 0.056. Conclusion: Simultaneous pharmacokinetic study in rat plasma and kidney is an effective method to investigate drug kidney penetration. RH can penetrate rat's kidney after administration to exert its effect.

Objective To elucidate the immunologic mechanism and clinical effect of high intensity focused ultrasound (HIFU) combined with dendritic cell and cytokine induced killer cell (DC-CIK) immunotherapy on patients with pancreatic cancer.Methods Seventy-two pancreatic cancer patients were divided randomly into 2 equal groups,one treated with HIFU only the other treated with HIFU and DC-CIK immunotherapy.Ultrasound imaging and a variety of immunological indexes were recorded before and after treatment and the clinical effects in the two groups were compared.Moreover,autogenous tumor cells were isolated from the combination therapy group and the killing activity of DC-CIK which loaded tumor antigen processed by HIFU on autogenous tumor cells was observed.Results Tumor antigen processed by HIFU can improve the killing activity of DC-CIK on autogenous tumor cells.After treatment,the immunological indexes,of all patients were better than before treatment.(58.26 ± 17.97 versus 52.15 ± 14.22 pg/ml with IL-12 22.14 ± 6.39 versus 17.36 ± 5.73 ng/ml with HSP70 and 0.94 ± O.34 versus 1.32 ± O.61 ng/ml with TGF-β,P ＜ 0.05 ) ; The combination group was significantly better than the HIFU group with regard to the average scores of quality of life (75.89 ± 19.65 versus 67.22 ± 16.34,P＜0.05),pain (3.15 ±0.82 versus 3.59 ± 1.04,P ＜0.05),tumor markers (107.55 ±27.58 versus 123.63 ±34.12 U/ml) and survival time (18.92±6.47 versus 13.36 ±5.78 mos).Conclusion HIFU can improve the immunologic status and anti-tumor response in patients with pancreatic cancer.HIFU combined with DC-CIK has good synergistic therapeutic effect for treating pancreatic cancer.

Objective To explore the immune effects of high intensity focused ultrasound (HIFU) in the treatment of pancreatic carcinoma and to investigate the imaging methods to evaluate HIFU's efficacy.Methods A total of 32 patients with pancreatic carcinoma treated by HIFU were enrolled.The freeze-thaw antigen was prepared by freezing and thawing the cancer cells.HIFU antigen was prepared by cancer cells sonicated by HIFU.The killing effects of no antigen activated dendritic cells (DC) induced T lymphocyte (DC-T),freeze-thaw antigen activated DC induced T lymphocyte (freeze-thaw antigen-DC-T) and HIFU activated DC induced T lymphocyte (HIFUantigen-DC-T) in autologous pancreatic cancer cells were detected by lactic dehydrogenase kit.The changes of immune indexes [heat shock protein 70 (HSP70),T helper lymphocyte Thl/Th2 and transforming growth factor-β (TGF-β)] before and after H IFU treatment were determined by enzymelinked immunosorbnent assay (ELISA) method.The changes of clinical efficacy indexes [visual analogue scale (VAS),performance status (PS) and carbohydrate antigen (CA) 19-9] before and after HIFU treatment were compared.The instant and recent (two months) efficacy of HIFU treatment were evaluated by contrast enhanced ultrasonograph (CEUS) and computed tomography (CT).The line q test was performed for comparision between groups.t-test was applied for comparision before and after treatment.Results Compared with freeze-thaw antigen,the killing effect of HIFU antigen-DC-T in autologous pancreatic cancer cells was higher (40.24％ ± 10.56％ vs 46.93％±13.26％,q=3.44,P＜0.05).HSP70 [(17.31±4.75) ng/mlvs (22.84±5.56) ng/ml],Th1/Th2 (1.24±0.36 vs 1.47±0.31),TGF-β [(1.39±0.41) ng/ml vs (1.04±0.38) ng/ml],VAS (3.97±1.32 vs 3.26±1.18),PS (2.76± 1.02 vs 2.21±0.86) and CA19-9 level[(135.39±37.45) U/ml vs (114.82±30.51) U/ml] improved after HIFU treatment compared with those before treatment (t=4.278,2.739,3.542,2.268,2.332 and 2.409,allP＜0.05).CEUS and CT showed that blood supply and the volume of the tumors reduced after HIFU treatment.Conclusions HIFU is effective in treating pancreatic carcinoma,improving immune status of patients and enhancing antitumor response.CEUS can real-time evaluate the efficacy of HIFU treatment.

Adenocarcinoma ; metabolism ; pathology ; surgery ; Adenocarcinoma, Papillary ; metabolism ; pathology ; surgery ; Antigens, CD ; metabolism ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; surgery ; Endoglin ; Female ; Humans ; Male ; Microvessels ; pathology ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; Receptors, Cell Surface ; metabolism ; SOX9 Transcription Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Survival Rate

Objective The role of long non-coding RNA Linc00467 in human lung adenocarcinoma is not yet clear.This study was to investigate the expression of long non-coding RNA Linc00467 in human lung adenocarcinoma, its clinical significance, and the effects of Linc00467 on the functions of the tumor and endothelial cells in vitro.Methods Lung adenocarcinoma tissue and normal tissue surrounding the malignance were obtained from 60 patients with pathologically proved stage I-Ⅲa lung adenocarcinoma.Human umbilical vein endothelial cells (HUVECs) were transfected with the over-expressed plasmid pccl-Linc00467 (HUVEC experimental group) or the empty vector pccl (HUVEC control group), A549 cells with Linc00467-siRNA (A549 experimental group) or negative siRNA (A549 control group), and H1299 cells, too, with Linc00467-siRNA (H1299 experimental group) or negative siRNA (H1299 control group).The expression level of Linc00467 in the lung adenocarcinoma tissue was detected by qRT-PCR with an analysis of its correlation with the clinicopathological characteristics of the patients;the influence of Linc00467 on the proliferation of the A549, H1299 and HUVEC cells was assayed with CCK-8;and the role of Linc00467 in the angiogenesis of the HUVECs was assessed by fibrin bead sprouting assay.Results The expression of Linc00467 in the lung adenocarcinoma tissue was 2.72±1.31 times as high as that in the normal lung tissue (P<0.01), and those in the A549 and H1299 cells were 3.45±0.25 and 3.22±0.33 times as high as those in the human bronchial epithelial (HBE) cells (P<0.01).The expression level of Linc00467 was significantly correlated with the tumor size and vascular invasion (P<0.05).After transfection of Linc00467-siRNA, the expressions of Linc00467 in the A549 and H1299 experimental groups were down-regulated by 72% and 68% as compared with those in the A549 and H1299 control groups (P<0.01).The number of living cells was remarkably decreased in the A549 experimental group in comparison with the A549 control at 48 h (1.29±0.07 vs 1.51±0.09), 72 h (1.53±0.15 vs 2.13±0.11), and 96 h after culturing (1.98±0.18 vs 3.02±0.12), and so was it in the H1299 experimental versus the H1299 control group, but markedly increased in the HUVEC experimental versus the HUVEC control group (P<0.05).At 5 days, HUVEC experimental group, as compared with the HUVEC control, showed a significantly increased number of newly formed vascular branches (7.36 vs 4.25/superbead, P<0.01) and relative length of the blood vessels (3.12 vs 1, P<0.01).Conclusion Linc00467 promotes tumor cell proliferation and angiogenesis and is highly expressed in the lung adenocarcinoma tissue, which is correlated with the tumor size and vascular invasion and suggests that Linc00467 could be a potential biomarker and therapeutic target.

Objective To explore the clinical features,diagnosi s and prognosis of incontinentia pigmenti.Methods Analyzing and summarizing the clinical characteristic, diagnosis and prognosis of neonatal incontinentia pigmenti in 6 neonatal infants that were hospita- lized in our department during the period from January 1 998 to December 2003 were studied,and some relevant literature were reviewed. Results 1.Three of 6 infants were male which was unusual;2.Four infants had typical skin lesions at birth and 1 case at 6 days old.Four cases had typical 3 stages o f skin lesions including the erythematous and vesicular inflammatory stage,verr ucous lesions and hyperkeratosis stage,macular hyperpigmentation stage,but the re was overlap;3.Four infants were complicated by central nervous system involv ement (two cases presented mental retardation,2 infants were temporary damage). Two cases were complicated by ocular manifestations ( one case had optical nerve atrophy and blind in left eye,the other had severe bilateral retinal lesions); 4.On specific examination 5 infants were diagnosed by skin biopsy.Gene analysis was made in 1 case,but we didn′t find the mutations of NEMO. Conclusions Incontinentia pigmenti is a rare X-linked dominant multisystem disease.It may be misdiagnosed in the initial stages.Except typical clinical features,skin biops y and gene analysis are main evidence for diagnosis.Early detection and interven tion are important for prognosis. J Appl Clin Pediatr,2005,20(2):123-125

Objective To improve the recognition of intramedullary spinal abscess by a case of congenital dermal sinus with intramedullary spinal abscess and reduco the incidence of congenital dermal sinus with intramedullary spinal abscess.Methods Clinical,laboratory data and image of a confirmed case about one infant of congenital dermal sinus with multiple intramedullary spinal abscess were investigated,the related literature was reviewed.Results In this case,when the infant with congential dermal sinus had infection,he failed to gain antibiotic therapy, timely surgical treatment,his infection had diffused, and multiple intramedullary spinal abscess flared up.Conclusions Intramedullary spinal abscess is a rare disease.If treatment is delayed, the prognosis is poor and the mortality rate is high.MRI is the ideal investigation for diagnosis.Intramedullary spinal abscess can happen subsequent to congenital dermal sinus with infection, and cause neurological sequela. So an infant with congenital dermal sinus should be offered to avoid complication caused by infection.

OBJECTIVETo analyze the relationship between clinical characteristics and prognosis of neonatal cerebral infarction and to draw attention to the disease to improve the long-term outcome through early diagnosis and intervention.

METHODSThe clinical characteristics of 6 confirmed cases were summarized. Perinatal conditions and other factors were analyzed for possible causes of the disease. The survived patients were followed-up for 6-8 months.

RESULTSThe authors diagnosed 6 cases of neonatal cerebral infarction in one year, which accounted for 0.6% (6/969) of all the in-patients in the same time period. Among them 3 cases were confirmed as cerebrovascular malformations by magnetic resonance angiography (MRA), In 1 case the infarction was due to severe bilateral intraventricular hemorrhage, and in another case the disease was related to comprehensive factors such as prematurity, maternal pregnancy induced hypertension and respiratory failure secondary to bronchopulmonary dysplasia (BPD), and in 1 case the cause was undetermined. Four out of the 6 patients presented with varied forms of convulsions, which became the second leading cause for all the neonatal convulsive events (20%). None of the patients had localized neurological signs in the early course except for abnormal muscular tone of some extent. Cerebral ultrasound scanning in 5 out of 6 cases showed positive results. The diffusion-weighted magnetic resonance imaging (DW-MRI) was highly valuable for early confirmative diagnosis. Only one case was found normal within one year of follow-up and all the other 5 cases had unfavorable prognoses of varied severity.

CONCLUSIONNeonatal cerebral infarction is not a rare condition and should be considered as one of the important causes for neonatal convulsion. Imaging study is the main technique for diagnosis. The prognoses were poor for those cases for whom early diagnosis and treatment can not be made or those with widespread cerebral lesions.

Brain ; blood supply ; pathology ; Cerebral Hemorrhage ; complications ; Cerebral Infarction ; diagnosis ; etiology ; Follow-Up Studies ; Humans ; Infant, Newborn ; Magnetic Resonance Angiography ; Male ; Prognosis ; Seizures ; etiology

Copy number aberrations (CNAs) in chromosome arm 8q have been associated with unfavorable clinical outcomes of several cancers and progressive tumor characteristics of hepatocellular carcinoma (HCC).This study was to identify correlation of CNAs in 8q with clinical outcomes of HCC patients,and further screen for differentially expressed genes in outcome-related CNAs.Array comparative genomic hybridization and expression arrays were performed to detect CNAs and expression levels,respectively.The correlations between CNAs in 8q and outcomes were analyzed in 66 patients,with a median follow-up time of 45.0 months (range,2.6-108.6 months).One hundred and nine cases were further evaluated to identify differentially expressed genes in the potential outcome-related CNAs.Copy number gain in 8q was observed in 22 (33.3 ％) of the 66 HCC cases.The most recurrent gains (with frequencies ＞20％) were 8q13.3-21.3,8q21.3-23.3,8q23.3-24.13,8q24.13-24.3,and 8q24.3.Survival analysis showed that 8q24.13-24.3 gain was significantly associated with reduced overall survival (P=0.010).Multivariate Cox analysis identified 8q24.13-24.3 gain as an independent prognostic factor for poor overall survival (HR=2.47;95％CI=1.16-5.26;P=0.019).A panel of 17 genes within the 8q24.13-24.3 region,including ATAD2,SQLE,PVT1,ASAP1,and NDRG1 were significantly upregulated in HCCs with 8q24.13-24.3 gain compared to those without.These results suggest that copy number gain at 8q24.13-24.3 is an unfavorable prognostic marker for HCC patients,and the potential oncogenes ATAD2,SQLE,PVT1,ASAP1,and NDRG1 within the regional gain,may contribute coordinately to the 8q24.13-24.3 gain-related poor prognosis.

AIM:To investigate the stemness of mouse triple-negative breast cancer (TNBC) 4T1 cells induced by doxorubicin (DOX) and the underlying mechanism.METHODS:The 4T1 cells and MDA-MB-468 cells were treated with DOX at different concentrations (0, 0.05, 0.1 and 0.5μmol/L) for 24 h, and the shape and viability of the cells were observed.The concentration of DOX at 0.1μmol/L was chosen as the optimal concentration for the following experiments.The 4T1 cells and MDA-MB-468 cells resistant to DOX were established by continuous stimulation with DOX for 4 weeks, and named as 4T1-DOX and MDA-MB-468-DOX.Sphere formation assay was used to detect the stemness of 4T1cells and MDA-MB-468 cells.The expression of CD133 was observed by immunofluorescence staining.The expression of CD44 was analyzed by flow cytometry.The protein levels of Stat3, phosphorylated Stat3 (p-Stat3) and Oct-4 were determined by Western blot.RESULTS:The sphere formation ability of the 4T1-DOX cells was stronger than that of the 4T1control cells.The 4T1-DOX cells expressed high levels of the stemness markers CD133 and CD44 as compared with the 4 T1 cells (P<0.05).Furthermore, the 4T1-DOX cells exhibited enhanced activation of Stat3 (p-Stat3) and increased expression of Oct-4 (P<0.05) , while the expression of total Stat3 had no obvious variation.In addition, when activation of Stat3 was inhibited by WP1066, the protein levels of p-Stat3, Oct-4 and CD44 were down-regulated (P<0.05).Furthermore, inhibition of Stat3 phosphorylation reduced the sphere formation ability of the 4T1-DOX cells (P<0.05).CONCLUSION:DOX induces the stemness of mouse TNBC 4T1 cells through Stat3-Oct-4 signaling pathway.