Objective To investigate the effects of hypoxia-inducible factor (HIF)-1α on glycolysis of human esophageal squamous carcinoma cells and the possible mechanism.Methods TE13 and Eca109 cells were cultured under normal oxygen (20％O2) and hypoxia (1％O2) conditions.The hypoxia was duration 6 hours,12 hours,24 hours and 48 hours.HIF-1α gene was stable silented by RNA interference method and TE13/small interfering HIF cells and Eca109/siHIF cells were obtained.The cell culture condition and time was same as TE13 and Eca109 cells.The changes of HIF-1α expression were detected by Western-blot.The changes of lactic acid concentration in cell culture supernatant were determined by Spectrophotometry.The changes of glucose transporter-1 (GLUT-1) and lactic dehydrogenase A (LDHA) expression at mRNA level were examined by realtime polymerase chain reaction.The changes of GLUT-1 and LDHA expression at protein level were tested by Western blot.Using t or t' test to analyze the effects of hypoxia duration on HIF-1αexpression at protein level.One-way ANOVA was applied for the difference analysis between the groups.Results In TE13 and Eca109 cells,the HIF-1α expression significantly increased under hypoxia condition and reached the peak at 12 hour (t=6.11,8.31; both P＜0.05).The lactic acid secretion of TE13/siHIF cells and Eca109/siHIF cells was (1.24±0.33) and (1.28±0.37) mmol/L,which significantly decreased when compared with TE13 and Eca109 cells [(3.25±1.34) and (4.91±1.69) mmol/L,t=2.53,3.59,both P＜0.05].The lactic acid secretion of TE13 and Eca109 cells significantly increased after hypoxia [(6.48±1.73) and (8.02± 1.95) mmol/L,t=2.715,2.050,both P＜0.05].There was no significant lactic acid secretion in TE13/siHIF cells and Eca109/siHIF cells after hypoxia (P ＞ 0.05).The expressions of GLUT-1 and LDHA at mRNA level were significantly suppressed in TE13/siHIF cells and Eca109/siHIF cells (normal oxygen:t=6.98,3.92,7.25,3.67,all P＜0.05).The expression of GLUT-1 at protein level remarkably weaked (normal oxygen:t=4.57、16.56,hypoxia:t=6.19、6.09,all P＜0.05),while the expression of LDHA at protein level slightly decreased (P＞0.05).Conclusions The level of glycolysis can be lowered by suppression HIF-1α expression in human esophageal squamous carcinoma cells.The pathway may be involved in the suppression of GLUT-1 and LDHA expression.Except for HIF-1α,there may be other regulating factors in LDHA protein expression at same time.

Objective To analyze the differentially expressed genes in esophageal squamons cell carcinoma (ESCC), para-cancerous tissue (PCT) and normal esophagus tissue (NET) using oligomicroarray and to identify the target genes related to the development and progression of esophageal carcinoma. Methods The total RNAs isolated from ESCC, PCT or NET using one step Trizol method were purified and reversely transcribed into cRNAs. The cRNAs were then fluorescence labeled and hybridized with Agilent oligomicroarray (21 074 probes). The fluorescence intensity features were detected by Agilent scanner and quantified by feature extraction software. The selected candidate genes were confirmed by real time real time fluorescent quantitative RT-PCR immunohistochemistry andWestern blotting.Results ① The oligomicroarray demonstrated that there were 38 up-regulated genes and 61 down-regulated genes. ② The real time fluorescent quantitative RT-PCR revealed that five genes (CTHRC1, INHBA, SPP1 ,LUM, HRK)were more differentially expressed in up-regulated genes. Of which, CTHRC1 displayed more disparity.③ Immunohistochemistry examination showed that the higher expression of CTHRC1 (56.5 %, 26/46) was observed in ESCC. There was significantly difference in expression of CTHRC1 between patients with or without lymph node metastasis (P<0.05). ④ CTHRC1 protein was expressed in both TE-13 and Eca-109 cell lines. Conclusion CTHRC1 is probably one of the most significant biomolecules in ESCC.

Rhynchophylla is a Chinese herb commonly used in clinical practice. It's also the primary herb of some famous Chinese herbal compound such as Tianma Gouteng decoction, and Lingyang Gouteng decoction. According the record from many previous materia medica literatures, rhynchophylla should be added later during decoction. Pharmaceutical research showed that rhynchophylla alkaloids were not stable. Which has resulted in many problems in the research and its application. For example, there was not a quantitative determination method in "Chinese Pharmacopoeia" of past and present versions, which seriously impacted its quality control and product application. Firstly, records from previous materia medica literatures and "Chinese Pharmacopoeia" were systematically sorted based on the chemical stability of rhynchophylla. Secondly, pharmaceutical research including chemical compositions and their stability, pharmacological effects, extraction process and quality analysis, was reviewed after reference of literatures published at home and abroad in recent decades. Positive reference and evidence for further research and development of rhynchophylla will be provided in the article.
Animals ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Quality Control ; Uncaria ; chemistry

OBJECTIVEThe demographic characteristics of acupuncture outpatient in China is investigated so as to provide clinical evidence for the establishment of acupuncture indication.

METHODSClinical epidemiological table was adopted to do cluster multistage random sampling in 36 hospitals of 31 provinces (municipalities and autonomous regions) in 2008. The composition ratios of all indices were analyzed with SPSS 13.0 software. And trend analysis was carried out on the patient's demographic characteristics and the age distribution of clinical indications.

RESULTSIn the year 2008, 31 858 cases of out patients from 36 hospitals of 6 administrative regions were collected. There were totally 14 411 male patients (45.2%) and 17 447 (54.8%) female. The gender ratio of male and female was 82: 100. The average age of the patients was (50.6 +/- 17.7) years old. The number of patients over 45 years old was more obvious than groups of other ages (P<0.05). Age differences can be found in those 6 regions (P<0.05). Clinical symptoms varied according to different ages of the patients. Different characteristics of age distribution of acupuncture indications were held to exist (P=0.000).

CONCLUSIONAt the present, differences of gender, age and region can be found in out patients of acupuncture clinic. And the result shows that the number of female patients is more than male, with a large proportion of patients over 45 years old. Differences of age distribution characteristics of acupuncture indications still exist, which is considered to have the relation with the dominate indications of acupuncture and aging of the population.

Acupuncture Therapy ; utilization ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; Demography ; Female ; Humans ; Infant ; Male ; Middle Aged ; Outpatients ; statistics & numerical data ; Young Adult

Objective The aim of this study was to investigate the distribution and clinical significance of CD8+ T cells in bladder cancer tissues in situ.Methods Immunohistochemistry were used to examine the distribution of CD8+ T cells in bladder cancer tissues,which were obtained from January 2003 to December 2009 from 302 patients.Among all the patients,262 were male while 40 were female;mean age is 60 years;tumor size ≤ 3 cm was in 235 and tumor size ＞ 3 cm was in 67;Unifocal tumor was in 214 and multifocal tumors were in 88.Amount of tumor stage Ta-T1 was 212 and T2-T4 was 90.Sixteen patients have lymph node metastasis.Histological low grade was diagnosed in 175 and histological high grade was diagnosed in 127.According to the differences between anatomic structure and cellular composition,bladder tumor tissues can be classified to two localization patterns:(1) intratumoral regions,defined as tumor cell nests;(2) stromal regions,defined as stromal areas that lack direct contact with tumor cells.Therefore,we divided 302 bladder cancer patients into two groups based on the median frequency of intratumoral CD8+ T cells (median,3/× 400 high resolution) and stromal CD8+ T cells (median,37/× 400 high resolution),respectively.x2 analysis was used to evaluated the correlation between CD8+ T cell density and clinicalpathological variables.Kaplan-Meier analysis and Cox proportional hazards regression models were applied to estimate overall survival (OS).Results CD8+ T cells were predominantly located in the intratumoral regions (mean,14 ± 2/× 400 high resolution) rather than in associated stromal regions (mean,50 ± 3/× 400 high resolution,P ＜ 0.05).The density of intratumoral CD8+ T cells was inversely associated with age (P =0.026),tumor size (P ＜ 0.05) and tumor stage (P ＜ 0.05),and could represent a favorable prognostic predictor of OS (HR =0.427,P =0.003).However,the density of stromal CD8+ T cells was positively associated with age (P =0.004) and histological grade (P ＜ 0.01),and could represent an adverse prognostic predictor of OS (HR =2.206,P =0.009).Conclusions Our findings suggest that intratumoral/ stromal CD8+ T cells could potentially serve as favorable/ adverse prognostic markers for bladder cancer patients,respectively.

Objective To investigate the changes of hypoxia-inducible factor(HIF)-1α and hexokinase-Ⅱ(HK-Ⅱ)expression in human esophageal squamous cell carcinoma and its effect in glycolysis.Methods TE13 cells and Eca109 cells were cultured under hypoxic condition(1 ％O2)for different hypoxic time(6,12,24 and 48 hours).Cells cultured under normal oxygen condition(20％O2)were set as control.The changes of HIF-1α and HK-Ⅱ expressions at protein level were detected by Western blot.HIF-1α genes were specifically silenced with RNA interference technology(RNAi),and then the changes of HIF-1α and HK-Ⅱ expression were determined by realtime PCR and Western blot.Under normal oxygen and hypoxic condition,the changes of lactic acid concentration in cell culture medium were detected by spectrophotometric method.Results Under hypoxic condition,the expression of HIF-1α and HK-Ⅱ gradually increased as hypoxic time extended(P＜0.05),reached a peak at 12h and then gradually decreased as time extended.Compared with that under normal oxygen condition,the expression of HK-Ⅱ in TE13 cells and Eca109 cells significantly increased under hypoxic condition(P＜0.05),which was more significant after 12 hours hypoxia.The result of realtime PCR indicated that under normal oxygen condition the expression of HIF-1α at RNA level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference(P＜0.05).The expression of HK-Ⅱ at RNA level was consistent with the result of HIF-1α.Under normal and hypoxia condition,the expression of HK-Ⅱ at protein level in TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).The lactic acid secretion of TE13 cells and Eca109 cells under hypoxia condition(14.707 ± 3.594 and 15.062 ±3.901)was higher than that under normal oxygen condition(6.070±1.839 and 6.891±1.592,P＜0.05).The lactic acid secretion of TE13/shRNA cells and Eca109/shRNA cells significantly decreased compared with TE13 cells and Eca109 cells without interference,and the difference was statistic significant(P＜0.05).Conclusion The expressions of HIF-1α and HK-Ⅱ in human esophageal squamous cell carcinoma significantly increased under hypoxia conditions.The expression of HK-Ⅱ is closely correlated with lactic acid concentration and HIF-1α expression.HIF-1α may affect cell glycolysis through HK-Ⅱ.

Objective To investigate the inhibitory effect of blocking PI3K/AKT pathway by wortmannin on hypoxia-inducible factor 1α (HIF-1α) and the effect on the expression of glycolysis associated genes in human esophageal carcinoma cell lines TE1 and TE13,and to analyze the relation between PI3K/AKT-HIF-1α pathway and glycolysis in esophageal carcinoma cells. Methods Esophageal carcinoma cell lines TE1 and TE13 pretreated with wortmannin (2 μmol/L) were incubated under normoxic and hypoxic conditions.And each cell line was divided into four groups.The expression of HIF-1α and glycolysis associated genes GLUT-1,LDHA and HK-Ⅱ at protein level were measured by.Western blot.The expression of HIF-1α,GLUT-1,LDHA and HK-Ⅱ at mRNA level was determined by real-time PCR. The activities of LDH and HK-Ⅱ and lactic acid (LA)concentration in the culture supernatant were tested with spectrophotometer method.Results Under normoxic condition,HIF-1α was expressed in TE1 cells and the expression of HIF-1α was inhibited by wortmannin (2 μmol/L),the most significant inhibitory effect was at 12 hours,therefore 12 hours was selected for the subsequent hypoxia experiment.Compared with untreated cells,the expression of HIF-1α、HK-Ⅱ 、GLUT-1、LDH-A at protein level significantly decreased in TE1 and TE13 cells after pretreated with wortmannin (P ＜ 0.05),and the expression of HIF-1α、HK-Ⅱ at mRNA level significantly decreased (P＜ 0.05).Under normoxic and hypoxic conditions,the HK-Ⅱ and LDH activities in TE1 and Te13 esophageal carcinoma cells significantly decreased after treated with wortmannin compared with untreated cells (P＜0.05).Under hypoxia condition,the enzyme activity increased in untreated cells (P＜ 0.05). Under normoxic and hyp0xic conditions,the lactic acid concentration in the culture supernatant obviously decreased in cells treated with wortmannin compared with untreated cells (P＜ 0.05). Under hypoxia condition,lactic acid concentration increased in wortmannin treated cells (P ＜ 0.05). Conclusions Under normoxic and hypoxic conditions,wortmannin decrease lactic acid concentration through inhibiting the expression of HIF-1α and glycolysis associated genes, which indicate PI3K/AKT-HIF-1α pathway was closely related to glycolysis in esophageal carcinoma cells.

Objective To investigate the imaging features of Ewing's sarcoma and primitive neuroectodermal tumor (ES and PNET) in sacrum.Methods Imaging data were retrospectively collected and reviewed in 17 patients with ES and PNET confirmed by pathology.X-ray was performed in all patients,CT scans in 13 patients,and MR scans in 10 patients.Results All lesions were detected solitary and lateral.X-ray and CT features were lytic bone destruction in 12 cases,2 of them with bone sclerosis; one patient showed diffuse sclerosis.Soft tissue mass in pelvic cavity can be seen in 11 cases,9 of them with adjacent large blood vessels invasive and 4 patients with sacroiliac joints invasive.On post-contrast images,mild enhancement was found in 8 cases,and 5 patients with marked enhancement.MR images showed intermediate signal intensity on T1WI in 8 cases and heterogeneous moderate to high signal intensity on T2WI/fat suppression in 10 cases.DWI showed high signal intensity in 4 cases.Unlike CT scan,contrastenhanced MR images showed significant enhancement in 7 cases.Conclusions Ewing's sarcoma and primitive neuroectodermal tumor (ES and PNET) in sacrum have some specific imaging features.Imaging examination could provide reliable evidence for clinical diagnosis.

Colitis, Ulcerative ; complications ; drug therapy ; Humans ; Male ; Middle Aged ; Psoriasis ; complications ; drug therapy ; Pyoderma Gangrenosum ; drug therapy ; etiology

Animals ; Cyclin-Dependent Kinase 5 ; metabolism ; Ethanol ; adverse effects ; Female ; Hippocampus ; drug effects ; metabolism ; Learning ; drug effects ; Male ; Memory ; drug effects ; Pregnancy ; Prenatal Exposure Delayed Effects ; metabolism ; physiopathology ; Rats