Objective To investigate the role of cancer stem cells in radiation resistance of esophageal cancer and its molecular mechanism, and to provide a theoretical basis for radiotherapy for esophageal cancer.Methods Esophageal cancer cell line TE1 was treated with 8 Gy of radiation. Esophageal cancer cell line with resistance to radiation, TE1-res, was established and screened.Cell counting was used to evaluate cell proliferation.Flow cytometry was used to determine the expression of CD44 (high) CD24(-) CD133(+) and apoptosis in cells.The colony formation assay was used to determine the colony-forming rate and cell survival curve.Bisulfite sequencing PCR was used to determine the methylation status of cancer suppressor genes.Comparison of the data was made by group t test or analysis of variance. Results Compared with TE1 cells, TE1-res cells had significantly enhanced proliferation, a significantly higher proportion of CD44( high) CD24(-) CD133(+) cells, and significantly enhanced resistance to apoptosis (mean value 20.84×105 vs.4.46×105/day, P=0.008;(38.0±2.9)%vs.(10.1±1.3)%, P=0.001;mean value 33.23% vs.10.50%, P=0.003 ) .After treatment with 8 Gy of radiation, TE1-res cells had significantly higher colony-forming rate and D0 value than TE1 cells ((14.3±2.6)%vs.(0.9±0.3)%, P=0.011;3.28 vs.2.19 Gy, P=0.125 ) .Moreover, the promoter methylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B was significantly enhanced in TE1-res cells than in TE1 cells ((89.7±4.9)%vs.(5.0±0.5)%, P=0.001;(92.3±4.7)%vs.(10.4±0.7)%, P=0.001;(90.7±3.7)%vs.(7.9±0.4)%, P=0.001;(83.4±5.7)%vs.(17.2±1.2)%, P=0.002;(90.2±
6.7)%vs.(4.4±1.2)%, P=0.002).Conclusions Cancer stem cells play an important role in radiation resistance of esophageal cancer. The resistance to radiation is closely associated with promoter hypermethylation in cancer suppressor genes including SPINT2, CDKN1B, DKK1, TP53, and PPP2R1B.

Objective To establish a radioresistant esophageal squamous cancer cell line,and to identify the radioresistant genes and mechanisms.Methods The radioresistant KYSE410-res cell line was established by repeated exposure of cell line KYSE410 to radiation.The proliferation and apoptosis of esophageal squamous cancer cells were evaluated before and after radiation.The changes in gene expression of the esophageal squamous cancer cells after radiation were determined by gene microarray and analyzed by group t test.The genes with significant difference in expression after radiation were validated.Results The KYSE410-res cells had significantly enhanced proliferation and anti-apoptosis than the KYSE410 cells (all P＜0.05).The result of gene microarray showed that compared with the KYSE410 cells,the KYSE410-res cells had the expression of 463 and 251 genes upregulated and downregulated by no less than 4 folds,respectively.Those genes with different expression levels after radiation were mainly responsible for cell proliferation,adhesion,signal transduction,angiogenesis,reactive oxygen metabolism,cell damage repair,and the MAPK/ERK signaling pathway.OAS2 and UBD were key proteins in the network.In the KYSE410-res cells,the expression of HLA-DQBI,MMP1,NCAM1,ZNF521,GPC6,SELENBP1,LCN15,and TFPI-2 genes measured by real-time PCR was consistent with that measured by gene microarray.Conclusions Abnormal activation of the MAPK/ERK signaling pathway,upregulated expression of OAS2 and UBD,downregulated expression of TFPI-2,and upregulated expression of MMPs may play a role in radioresistance of esophageal cancer cells.

Because of the high morbidity and mortality rate,cardio-cerebrovascular diseases,including cerebral embolism, cerebral hemorrhage,cerebral vasospasm and myocardial infarction,have become main diseases threatening human health. Tradition-al chinese medicine(TCM)holds that the basic pathogenesis is Qi imbalances,which could be improved by benefiting Qi and promot-ing blood circulation. The compatibility of Radix astragali and Radix salviae miltiorrhiae are particularly suitable for treating the car-dio-cerebrovascular system diseases by improving Qi deficiency and blood stasis. This paper focuses on the application of Radix astrag-ali,Radix s. miltiorrhiae and their compatibility for treating the cardio-cerebrovascular system diseases. Moreover,our research would offer valuable references for the development of new drugs related to the treatment of cardio-cerebrovascular system diseases.

Administration, Intravaginal ; Adult ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Microwaves ; therapeutic use ; Middle Aged ; Phytotherapy ; Uterine Cervical Erosion ; therapy

Objective To explore the characteristics of etiology,clinical,electroencephalogram(EEG) and prognosis of infantile spasms(IS) with focal seizures(FS).Methods The significance of age onset,seizure patterns and atteration,etiology,video-EEG(VEEG) and evolution of FS correlating to epileptic spasms(ES),which occurred in 12 cases with IS by means of clinical observation,cranial CT or MRI,VEEG monitory and follow up were investigated.Twelve cases were divided into group A,B,C according to the stages of FS occurring prece-ding,coinciding and following ES.Results Ten cases with IS were identified from focal cortical dysplasia,tuberous scleroses complex,temporal lobe cyst or scleroses and hypoxic-ischemic encephalopathy and others.Clinical manifestation presented FS at certain times during the course of the disease,and other characteristics of frequent attacks,asymmetric spasms or tonic spasms,some atypical seizures in the eyes and the head,motionless staring and focal motor seizures and other.The correlation of ES to FS occurred during one ictal episode as follows:FS→ES(6 cases),ES→FS→ES(1 case),ES→FS(3 cases).FS appeared more frequently,atypical,predominantly involving ocular,facial,oral movement or generalized convulsion,migrating or alternating seizures,associated with epileptic discharges of posterior parietal-occipital and parietal-temporal-occipital origins in group A and early period of group B;whereas in the late period of group B and group C,FS occurred less frequently and more stable,presenting complex partial seizures or secondarily generalized seizures originated mainly from frontal and surrounding areas in the lateralized or bilateral hemispheres.Regarding the natural evolution and development of IS,2 cases with early FS developed into IS;6 cases with IS evolved into Lennox-Gastaut syndrome or symtomatic generalized epilepsy,4 cases got into FS,1 case maintained in a peculiarly epileptic state with FS as well as ES until the operation at 4.5 years old,and the other case was not identified clearly.Conclusions IS coincided with FS is a special kind of aged-related FS associated with secondarily generalized seizures and epileptic encephalopathy.The multiple etiology,seizure patterns,ictal-interictal EEG,clinical evolution and prognosis of the disorder,indicate a complicated interaction of the immature cortico-subcortical abnormalities in the critical developmental period and thereafter,can be identified as a peculiar form of clinical epileptic syndrome.

A rapid LAMP detection method with primers designed on genus-specific region identified in the gyrA gene was established in this assay. All four Campylobacter jejuni from different sources were detected positive and fourteen non Campylobacter bacteria were negative, which shows excellent specificity of the primers. Compared with plate count and PCR method, the LAMP method and the PCR method had equal sensitivity, which were three orders of magnitude higher than plate count. In this assay, we also found out that the treatment of DNase could reduce the dead bacteria DNA effectively. The LAMP detection on chicken indicated relatively good result on detection of Campylobacter jejuni combining with treatment of DNase.

Objective To evaluate the effect of angiotensin Ⅱ (Ang Ⅱ ) on the change of nephrin phosphorylation both in Ang Ⅱ-infused rat model and cultured podocytes.Methods Thirty Wistar rats were subcutaneously embedded with osmotic minipumps and randomly divided into 3 groups according to receiving either Ang Ⅱ at a dose of 400 ng· kg-1· min-1 or Ang Ⅱ +telmisartan at a dose of 3 mg·kg-1 ·d-1,or normal saline as a control group.Blood pressure and 24-hour urinary albumin were measured at 0 d,7 d,14 d,21 d and 28 d of the experiment.Renal histomorphology was evaluated through electron microscopy.The concentrations of Ang Ⅱ both in blood plasma and kidney were detected by radioimmunoassay.In vitro,cultured murine podocytes were exposed to Ang Ⅱ (10-6 mol/L) pretreated with or without losartan (10-5 mol/L) for different time periods.Nephrin and its phosphorylation expression were analyzed by Western blotting.The distribution of F-actin was presented by FITC-phallodin labeling.The change in phenomenon of F-actin was evaluated by cortical F-actin score index (CFS).Results (1)Ang Ⅱ-infused rats exhibited increased Ang Ⅱ concentration,significant hypertension and marked albuminuria.(2)In Ang Ⅱ-infusion group,nephrin expression was decreased (P＜0.05).Ang Ⅱ-receiving rats displayed diminished phosphorylation of nephrin.(3)In vitro,the phosphorylation of nephrin was significantly reduced after Ang Ⅱ stimulation for 3-6 hours (P＜0.05).(4)Ang Ⅱ stimulatation resulted in irregularly arrangement of F-actin followed by the redistribution of F-actin to podocyte periphery and formation of F-actin ring,in which the CFS obviously increased compared to control (P＜0.05).Conclusions Phosphorylation of nephrin is important for the survival status of podocytes.Ang Ⅱ-induced nephrin dephosphorylation may be an important molecular mechanism for Ang Ⅱ-induced podocyte cytoskeleton rearrangement and foot process effacement.

Most Chinese medicine has good clinical efficacy and application. However, the material basis is vague for the lack of basic research, the value of Chinese medicine is hard to reflect for the low technology level and product quality is difficult to maintain for the quality control indicator selection is incorrect. Chinese medicine Jinfukang oral liquid is a typical product of Chinese medicines. Jinfukang oral liquid was selected as a model drug in this article. Based on the overall concept and systems theory of traditional Chinese medicine, the research idea and technology system for modern Chinese medicine secondary exploitation was formed. The system includes three parts, for the first, basic research to make clear the components structure and their action mechanism, for the second, technology upgraded to optimize process and improve the product quality, for the last, exploring the associated industry to form the industrial chain. The research ideas and technology system based on the material basis research and development of modern Chinese medicine, guided with component structure in Chinese medicine and aimed clinical needs. This research ideas and technology system provides strategies and methods for the development of modern Chinese medicine secondary exploitation.
Administration, Oral ; Chemistry, Pharmaceutical ; standards ; Drug Therapy ; standards ; Drugs, Chinese Herbal ; administration & dosage ; standards ; Humans ; Quality Control

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

Background Our pervious antibacterial studies on several traditional Chinese medicines have found that Patchouli oil from Pogostemon cablin had significant antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA),which has spread worldwide and infected innumerable people.In order to find the more active natural substances in Patchouli oil,one of the major components,Pogostone,was isolated and its antibacterial activity was evaluated in vitro and in vivo in this study.Methods In vitro test,Pogostone was screened for antimicrobial properties against 83 bacteria comprising 35 gram positive and 48 gram negative bacteria strains via the agar double dilution method.In vivo test,specific pathogen free (SPF) strain of both male and female white Kunming mice,weighing 18-22 g,were used to test the protective ability of Pogostone after being injected with the median lethal doses (MLDs) of the tested strains.Results In vitro test,Pogostone could inhibit both gram negative bacteria (0.098-1 600 μg/ml) and gram positive bacteria(0.098-800 μg/ml).For Corynebacterium xerosis and some Chryseobacterium indologenes,the minimum inhibitory concentration (MIC) values of Pogostone were extremely low (＜0.098 μg/ml).It was significant that Pogostone was also active against some drug-resistant bacteria like MRSA.Furthermore,Pogostone showed antibacterial activity in vivo against Escherichia coli (E.coli) and MRSA via intraperitoneal injection.Ninety percent of the mice infected with E.coil could be protected at the concentrations of 50 and 100 mg/kg,and 60％ of the mice at 25 mg/kg,while the rate of protection for the mice infected with MRSA was 60％ and 50％ at doses of 100 and 50 mg/kg,respectively.Conclusion Pogostone could be developed as a potential antibacterial agent for clinical therapy.