Objective To discuss the effects of high glucose on human mesangial cells (HMC)Wnt-β-catenin signaling pathway,and the intervention of rhein on it.Methods High concentration of glucose (30 mmol/L) combined with different concentrations of rhein were used to intervene cultured human mesangial cells.The activity of mesangial cell proliferation after all the interventions was examined by MTT measurement.Total Wnt、β-catenin RNA was detected By RT-PCR in normal mesangial cells and cells intervened by high glucose and Rhein.Results ①Inhibition effect to human mesangial cells:compared with NG group for 24 h,48 h and 72 h (OD values were 0.169± 0.051,0.228±0.074,0.285±0.075),human mesangial cells proliferation in HG group(OD values were 0.307± 0.074,0.507 ±0.038,0.711±0.075),HG+R1 group(OD value were 0.241± 0.027,0.334±0.015,0.499±0.063),HG+R2 group (OD value were 0.244±0.081,0.386±0.033,0.531±0.011),and HG+R3 group(OD value were 0.277±0.036,0.407± 0.057,0.594±0.042) were iucreased significantly (P＜0.05、P＜0.01) ; Compared with HG group for 24 h,48 h and 72 h,the HG+R1 group,the HG+R2 group and the HG+ R3 group showed a downward trend at 24 h,but not significantly; but the trend decreased significantly at 48 h and 72 h,and the performance expressed a time,concentration-dependent (P＜0.05,P＜0.01).② In the normal state,the mesangial cells expressed s certain amount of Wnt and β-catenin,when they were stimulated by high glucose,the expression of Wnt、β-catenin in mRNA increased (P＜0.05) ; while the expression of them was significantly reduced in high glucose-induced mesangial cells in HG+R1 group,HG+R2 group and HG+R3 group(P＜0.05).Conclusion Rhein may inhibit the proliferation of high glucose-induced mesangial cells through the Wnt and β-catenin gene expression.

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.

Chinese traditional patent medicine for promoting blood circulation and removing blood stasis(PBCRBS) originated from traditional Chinese medicine theory and had approved efficacy and safety standards. However, its compatibility regularity and anti-thrombotic mechanism is not clear. To analyze the compatibility regularity and anti-thrombotic mechanism of Chinese traditional patent medicine for PBCRBS, a statistical and bioinformatics analysis was carried out using traditional Chinese medicine inheritance support system (TICMISS, V2.0) and ingenuity pathway analysis (IPA). The compatibility regularity analysis shows that the most commonly used herb combinations are Danshen (Salvia miltiorrhiza Bge.), Chuanxiong (Ligusticum chuanxiong Hort.) and Honghua (Carthamustinctorius L.). The anti-thrombotic mechanism analysis reveals that 25 ingredients have an effect on 29 thrombosis related molecules which 23 molecules are related to inflammation response. Furthermore, there are 5 inflammation molecules (NOS2, PTGS2, IL6, TNF, IL1β) served as major targets. At the same time, Danshen, Chuangxiong and Honghua mainly used as sovereign herb or minister herb in the application of cardiovascular and cerebrovascular diseases. Therefore, Chinese traditional patent medicine for PBCRBS probably has an effect on anti-thrombotic activity through inhibiting the inflammatory response. In summary, the most commonly used herb combinations of Chinese traditional patent medicine for PBCRBS are Danshen, Chuanxiong and Honghua. Inhibiting inflammatory response, especially inflammation related molecules (NOS2, PTGS2, IL6, TNF and IL1β), is probably a new starting point to clarify the anti-thrombotic mechanism of Chinese patent medicine for PBCRBS.
Anti-Inflammatory Agents ; pharmacology ; Carthamus tinctorius ; Computational Biology ; Drugs, Chinese Herbal ; pharmacology ; Fibrinolytic Agents ; pharmacology ; Humans ; Inflammation ; drug therapy ; Medicine, Chinese Traditional

OBJECTIVETo explore the role of TBX3 gene in the pathogenesis of breast cancer.

METHODSThe total RNA of 51 fresh breast cancer tissues and the corresponding adjacent tissues were extracted and reverse transcribed into cDNA to detect the expression of TBX3 mRNA by real-time PCR. The correlation between TBX3 mRNA expression and the clinicopathologic parameters in relation to breast cancer metastasis was analyzed.

RESULTCompared to that in the adjacent tissues, the expression of TBX3 mRNA was markedly increased in breast cancer tissues. TBX3 mRNA expression was significantly higher in metastatic breast cancer than in non-metastatic tumors.

CONCLUSIONIncreased expression of TBX3 mRNA suggests the involvement of TBX3 in the pathogenesis and metastasis of breast cancer.

Breast Neoplasms ; etiology ; genetics ; pathology ; Female ; Humans ; Neoplasm Metastasis ; genetics ; RNA, Messenger ; genetics ; metabolism ; T-Box Domain Proteins ; genetics ; metabolism

OBJECTIVETo investigate the feasibility of continuous intraspinal ceftazidime administration for treatment of purulent meningitis due to Achromobacter infection.

METHODSA patient with established diagnosis of purulent meningitis due to Achromobacter infection was admitted, who failed to respond favorably to a 3-day ceftazidime treatment administered intravenously. Continuous intraspinal ceftazidime administration at the dose of 0.2 g/d was then attempted through a catheter placed in the cisterna magna in addition to intravenous ceftazidime for 3 days, which resulted in obvious relief of the symptoms. The catheter was subsequently withdrawn, and the patient received further treatment with additional intravenous ceftazidime for a week.

RESULTSThe symptoms of purulent meningitis was significantly improved after a 3-day continuous intraspinal ceftazidime administration, and the patient was eventually cured after completion of the treatment course. Intrathecal ceftazidime was also attempted previously but failed due to intolerance of pains in the legs. No relapse was observed in this case 3 months after the discharge.

CONCLUSIONContinuous intraspinal ceftazidime administration can be effective and safe for treatment of purulent meningitis.

Achromobacter ; Adult ; Anti-Bacterial Agents ; therapeutic use ; Catheters, Indwelling ; Ceftazidime ; therapeutic use ; Humans ; Injections, Spinal ; Male ; Meningitis, Bacterial ; drug therapy ; Treatment Outcome

OBJECTIVETo investigate the feasibility of real-time PCR-optimized oligonucleotide probe method for detection of KRAS mutations in lung and colorectal carcinomas, as compared with Sanger sequencing method.

METHODSGenomic DNA was extracted from formalin fixed paraffin embedded samples of 221 lung carcinomas and 131 colorectal carcinomas after tumor cell content assessment and macrodissection. Real-time PCR-optimized oligonucleotide probe method and Sanger sequencing were performed to detect KRAS gene mutations. The frequency of KRAS mutation, mutation types, and their concordance were analyzed.

RESULTSKRAS mutation was detected in 6.3% (14/221) and 4.5% (10/221) of 221 lung cancer samples by using real-time PCR-optimized oligonucleotide probe method and Sanger sequencing, respectively, while in 41.2% (54/131) and 40.5% (53/131) of 131 colorectal cancer samples, respectively. There was no significant correlation between KRAS gene mutations and patients' gender and age (P > 0.05). The positive rate of KRAS codon 12 mutation was significantly higher than that of KRAS codon 13 mutation (P < 0.05). The overall concordance between real-time PCR-optimized oligonucleotide probe method and Sanger sequencing for KRAS mutation detection was 97.4%.

CONCLUSIONReal-time PCR-optimized oligonucleotide probe method provides an alternative method with high consistency and sensitivity as compared to Sanger sequencing in gene mutation detection.

Codon ; Colorectal Neoplasms ; genetics ; DNA Mutational Analysis ; methods ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Mutation ; Oligonucleotide Probes ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins p21(ras) ; Real-Time Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; ras Proteins ; genetics

OBJECTIVETo investigate the mutation frequencies of KRAS, BRAF, PIK3CA and EGFR genes that were effective on the targeted therapies in colorectal carcinoma.

METHODSThe tissue specimens from 331 colorectal cancer patients were collected and subject to KRAS, BRAF, PIK3CA and EGFR mutation analysis. Paraffin-embedded tissue samples were obtained and macrodissection was performed to enrich the tumor cells for DNA extraction when necessary. PCR-based direct DNA sequencing was used to investigate the codons 12 and 13 in exon 2 of KRAS gene, exons 11 and 15 of BRAF gene, exons 9 and 20 of PIK3CA gene and exons 18-21 of EGFR gene.

RESULTSActivating mutations were detected in KRAS (44.1%, 137/311), BRAF (5.8%, 9/156), PIK3CA (2.6%, 4/156) and EGFR (1.3%, 2/156) in the study cohort of colorectal carcinoma cases. Among KRAS gene mutations, 81.0% (111/137) occurred in codon 12, with p.G12D as the most common variant (45.3%, 62/137); 19.0% (26/137) occurred in codon 13, with 38G > A (G13D) as the most common variant (17.5%, 24/137).

CONCLUSIONSThe KRAS mutation frequency is the highest among the four genes (KRAS, BRAF, PIK3CA and EGFT) tested in colorectal carcinoma. The presence of these gene mutations may provide therapeutic information for targeted therapy. Mutation analyses of BRAF and PIK3CA in addition to KRAS should be a part of the standard diagnostic algorithm for colorectal carcinoma patients.

Class I Phosphatidylinositol 3-Kinases ; Codon ; Colorectal Neoplasms ; genetics ; pathology ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Phosphatidylinositol 3-Kinases ; genetics ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins B-raf ; genetics ; Proto-Oncogene Proteins p21(ras) ; Receptor, Epidermal Growth Factor ; genetics ; ras Proteins ; genetics

BACKGROUNDY-27632 is a specific inhibitor of Rho-associated coiled kinase (ROCK) and has been shown to promote the survival and induce the differentiation of a variety of cells types. However, the effects of Y-27632 on adult human adipose tissue-derived stem cells (ADSCs) are unclear. This study aimed to investigate the effects of Y-27632 on the neuronal-like differentiation of ADSCs.

METHODSADSCs were isolated from women undergoing plastic surgery and cultured. ADSCs were treated with different doses of Y-27632 and observed morphological changes under microscope. The expression of nestin, neuron specific enolase (NSE) and microtubule-associated protein-2 (MAP-2) in ADSCs treated with Y-27632 was detected by immunocytochemistry and Western blotting analysis.

RESULTSY-27632 had the potency to induce neuronal-like differentiation in ADSCs in a dose-dependent manner. Moreover, the differentiation induced by Y-27632 was recovered upon drug withdraw. ADSCs treated with Y-27632 expressed neuronal markers such as NSE, MAP-2 and nestin while untreated ADSCs did not express these markers.

CONCLUSIONSelective ROCK inhibitor Y-27632 could potentiate the neuronal-like differentiation of ADSCs, suggesting that Y-27632 could be utilized to induce the differentiation of ADSCs to neurons and facilitate the clinical application of ADSCs in tissue engineering.

Adipose Tissue ; cytology ; Adult ; Amides ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Female ; Humans ; Neurons ; cytology ; Pyridines ; pharmacology ; Stem Cells ; cytology ; drug effects

BACKGROUNDBone morphogenetic protein-6 (BMP-6) is closely correlated with tumor differentiation and skeletal metastasis. Estrogen is considered as a stimulant for the initiation and promotion of breast cancer. Previous studies demonstrated that 17beta-estadiol (E2) can selectively increase the expression of BMP-6. This experiment is designed to detect the molecular mechanism of estrogen activating BMP-6 gene transcription in human estrogen receptor positive (ER+) breast cancer cell line MCF-7.

METHODSAfter the treatment of MCF-7 cells with E2 at different concentrations (10(-11) mol/L, 10(-9) mol/L, 10(-7) mol/L), the BMP-6 expression level was examined through real-time polymerase chain reaction. Through restriction enzyme digestion, human BMP-6 1.2 kb long promoter, BMP-6 0.7 kb long promoter was cloned into pGL-3 basic vector; after the treatment with 10(-7) mol/L E2, luciferase activities of the two promoters were detected. Site-directed mutagenesis was performed to obtain the mutant forms of estrogen response element half-site (1/2 ERE) element and Sp1 sites in the BMP-6 promoter, the activities of these mutant form promoters were detected following the methods mentioned above. Chromatin immunoprecipitation (ChIP) assay was also used to confirm the binding of estrogen receptor alpha (ERalpha) on BMP-6 promoter in the presence of E2.

RESULTSE2 dose dependently increased BMP-6 mRNA expression in human ER+ breast cancer cell line MCF-7. At a dose of 10(-7) mol/L E2, human BMP-6 1.2 kb promoter activity was increased by 90% compared with the control group treated with ethanol (P < 0.05). Both the 1/2 ERE response element mutant form and the Sp1 site mutant form of the BMP-6 promoter abolished the activation of the BMP-6 promoter's response to E2. Through ChIP assay, the binding of ERalpha on 1/2 ERE response element in BMP-6 promoter was further validated.

CONCLUSIONEstrogen induces BMP-6 expression in human ER+ breast cancer cell line MCF-7 through its receptor ERalpha binding on 1/2 ERE element in the BMP-6 promoter.

Bone Morphogenetic Protein 6 ; Bone Morphogenetic Proteins ; genetics ; Breast Neoplasms ; genetics ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; physiology ; Female ; Humans ; Parathyroid Hormone-Related Protein ; secretion ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects

OBJECTIVEThe National Institutes of Health (NIH) category IIIa chronic prostatitis syndromes (non bacterial chronic prostatitis) were common disorders but with few effective therapies. Alpha-blockers and bioflavonoids had recently been reported in randomized controlled trials to improve the symptom of these disorders in a significant proportion of men. The aim of this study was to confirm these findings in a prospective randomized, placebo-controlled trial.

METHODSForty-five men with category IIIa chronic non bacterial protatitis were randomized into three groups as follows: (1) placebo; (2) phenoxybenzamine-hydrochloride:10 mg two times a day for one month; (3) flavoxate HCI-neptumus: 200 mg three times a day for one month. The NIH chronic prostatitis symptom score was used to grade symptoms at the beginning and conclusion of the study.

RESULTSAll the patients in three groups completed the study except three dropout patients in placebo group because of sever symptoms. The three groups were similar in age, duration of symptoms and initial symptom score. Patients taking placebo had a mean improvement in NIH-CPSI from 21.85 to 19.55 (not significant), while the phenoxybenzamine-hydrochloride group had a mean improvement from 21.95 to 13.75 (P < 0.01), and those taking flavoxate HCI-neptumus had a mean improvement from 21.75 to 16.95 (P < 0.05). The decrease in NIH-CPSI was associated with significant improvement in patients' clinical manifestations.

CONCLUSIONTherapy with alpha-blockers was well tolerated with significant symptomatic improvement in most men having chronic non-bacterial chronic protatitis while the bioflavonoids group had no significant improvement. Mechanism of both medicines needs further study.

Adrenergic alpha-Antagonists ; administration & dosage ; therapeutic use ; Adult ; Chronic Disease ; Flavonoids ; administration & dosage ; therapeutic use ; Flavoxate ; therapeutic use ; Humans ; Male ; Parasympatholytics ; therapeutic use ; Prospective Studies ; Prostatitis ; drug therapy ; Treatment Outcome