Kidney injury and decreased chemosensitivity of tumor cells are obstacles with cisplatin (CDDP) chemotherapy. Down-regulation of the organic cation transporter 2 (OCT2) and multidrug resistance-associated protein 2 (MRP2) is a key means to alleviate CDDP-induced kidney injury and increase chemosensitivity. Astragaloside IV (AS IV) is obtained from the well-known traditional Chinese herb Astragalus membranaceus. This study explored the role of AS IV in preventing kidney injury and enhancing the antitumor effect of CDDP by suppressing OCT2 expression in kidney and MRP2 in tumors. This project was reviewed and approved by the Animal Ethics Committee of the First Hospital of Jilin University. The effects of AS IV on CDDP inhibition of tumor growth and promotion of apoptosis were assessed in Lewis lung tumor (LLC)-bearing mice by H&E and TUNEL staining. Kidney injury was assessed by serum biochemical parameters and H&E staining. We used Western blotting and immunohistochemistry assays to detect OCT2 and MRP2 expression in kidney and tumor. The concentration of CDDP in kidney and tumor was measured by HPLC-MS/MS. AS IV enhanced CDDP chemosensitivity by increasing tumor cell apoptosis and slowing tumor growth, and decreased kidney injury as evidenced by lower blood creatinine (Cr) and blood urea nitrogen (BUN). Co-administration of AS IV suppressed MRP2 overexpression induced by CDDP in tumor tissues and may be an important mechanism for enhancing CDDP chemosensitivity. Moreover, AS IV reduced CDDP-induced kidney injury in mice along with suppression of OCT2 expression in kidney. The concentration of CDDP was increased in tumor but decreased in kidney. In total, AS IV not only enhanced the antitumor effect of CDDP by suppressing MRP2 expression in tumor cells, but also decreased kidney injury induced by CDDP. The results provide new insight into the combined use of a chemotherapy drug and natural ingredients to treat cancer.

Objective To study the anti-tumor effects of humulon on arylamine N-acetyltransferase-1(NAT1)activity.Methods Employing HPLC,using PABA as substrate,in intact SGC-7901 cells and their cytoplasm,making PABA being acetylated to Ac-PABA by NAT1 as the activity of NAT1.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to study the expression of the NAT1 mRNA.Results The results show that humulon could inhibit the production of Ac-PABA in intact SGC-7901 cell and the cytoplasm,the production of Ac-PABA was gradually increased with the interaction time increasing.But comparing with corresponding negative control group's,the production of Ac-PABA was decreased evidently and the humulone could inhibit the expression of NAT1 mRNA.Conclusion Humulon could prevent the occurrence and deterioration of cancer.Its mechanisms can be attributed to its effect on decreasing the production of acetylation of carcinogenic aromatic amines,which is acetylated from aromatic amines,and inhibiting the NAT1 activity and expression of NAT1 mRNA.

Objective To explore the effects of humulon on kinetic parameters of N-acetyltransferase-1(NAT1) of human gastric cancer SGC-7901.Methods Employing HPLC,using para-aminobenzoic acid(PABA) as substrate,in intact SGC-7901 cells and their cytoplasm,taking the speed of PABA being acetylated to Ac-PABA by NAT1 as the rate of NAT1,using double reciprocal plot,taking the reciprocal of concentration of PABA and reaction rate of NAT1 as coordinates,regression equation was obtainied and the Michaelis constant(Km) and maximum reaction velocity(Vmax) were calculated.Results Study on enzyme kinetics demonstrated,as for intact SGC-7901 cells,Km and Vmax of control group were(3.910?0.087) ?mol/L and(0.306 0?0.006 7) pmol/L(1?106 cells),respectively,Km and Vmax of the humulon group were(3.830?0.123) ?mol/L and(0.275 0?0.005 8) pmol(1?106 cells),respectively.As for the cytoplasm of SGC-7901 cells,Km and Vmax of control group were(760.2?210.2) ?mol/L and(0.191 0?0.043 7) pmol/(mg?min),Km and Vmax of the humulon group were(449.0?72.9) ?mol/L and(0.094 0?0.010 4) pmol/(mg?min).Statistically,as for intact SGC-7901 cells or their cytoplasm,there was no difference of the Km between control group and humulon group,but there was remarkable difference of Vmax between control group and humulon group,P

OBJECTIVETo observe the therapeutic effect of moxibustion and exercise comprehensive scheme intervention for children with short stature of deficience of the kidney essence.

METHODSTwenty four cases of children in 12 to 14 years old were selected, 12 male and 12 female, they were treated with comprehensive therapy of exercise therapy and moxibustion. Running and jumping were selected as main exercise therapy, it became a suitable exercise amount when the heart rate reach to 150 to 170 times per minute, thrice each week, 35 to 45 minutes each time. After exercises they were treated with moxibustion, Qihai (CV 6), Guanyuan (CV 4), Zusanli (ST 36), Dazhu (BL 11), Xuanzhong (GB 39), Geshu (BL 17) etc. were selected. After treatment for half a year, the changes of the body height, body weight, bone age(BA), growth hormone (GH), testosterone (T) and estradiol (E2) were compared before and after treatment.

RESULTSThe body height and bone age of the boys and girls were significantly higher than those before treatment (all P<0.05), the growth of body height was more than 4 cm, the growth of bone age was more than 0.5 years old in half a year; the testosterone of all children was significantly increased (all P<0.05), and there were no significant differences in body weight, GH and E2 compared to those before treatment (all P>0.05).

CONCLUSIONMoxbustion and exercise comprehensive scheme can effectively improve the children with short stature of deficience of the kidney essence, the mechanism is related to the improving of the testosterone level.

Adolescent ; Body Height ; Child ; Estradiol ; metabolism ; Exercise Therapy ; Female ; Growth Disorders ; metabolism ; physiopathology ; therapy ; Human Growth Hormone ; metabolism ; Humans ; Kidney ; physiopathology ; Male ; Moxibustion ; Testosterone ; metabolism ; Treatment Outcome

OBJECTIVETo analyze the extent of myocardium and coronary artery lesion post atrioventricular ring radiofrequency catheter ablation with different tip catheters.

METHODSTwenty-one healthy dogs were randomly divided into 64 degrees C/50 W/100 s, 64 degrees C/100 W/100 s, 45 degrees C/45 W/100 s groups and ablated by 4 mm tip catheter, 8 mm tip catheter and irrigated tip catheter respectively. Left atrioventricular ring and right atrioventricular ring ablation were performed in all dogs. After ablation, myocardium lesion volume was calculated as 1/6pi x length x width x depth. Histological examinations were performed at the myocardium tissue at ablation sites.

RESULTSThe lesion depths post 8 mm tip catheter ablation (7.18 +/- 1.72) mm and irrigated tip catheter ablation (7.99 +/- 1.77) mm were similar and significantly deeper than that post 4 mm tip catheter ablation (4.54 +/- 1.38) mm, P < 0.01. Similar results were found in terms of lesion volume [(356.76 +/- 94.44) mm(3) post 8 mm tip catheter ablation, (391.69 +/- 109.54) mm(3) post irrigated tip catheter ablation and (191.34 +/- 74.52) mm(3) post 4 mm tip catheter ablation]. Five (5/42, 11.9%) transmural myocardium necrosis and 8 (8/42, 19%) coronary artery lesions were observed post ablations.

CONCLUSIONThe extents of post ablation myocardium and coronary artery lesion were significantly higher induced by 8 mm tip catheter and irrigate tip catheter compared those by 4 mm tip catheter.

Animals ; Cardiac Catheterization ; adverse effects ; Catheter Ablation ; adverse effects ; Coronary Vessels ; pathology ; Dogs ; Myocardium ; pathology

OBJECTIVETo investigate the relationship between RASSF1A gene expression and DNA methylation or histone modification in laryngeal carcinoma tissues.

METHODSChromatin immunoprecipitation (ChIP), methylation specific polymerase chain reaction (MSP) and realtime quantitative reverse transcription polymerase chain reaction (realtime RT-PCR) were used to analyze RASSF1A gene promoter region histone H3 lysine 9 methylation, H3 lysine 4 methylation, H3 lysine 9 acetylation, DNA methylation, and RASSF1A gene expression in laryngeal carcinoma tissue of 50 cases.

RESULTSDNA methylation rate of gene RASSF1A was 62% in 50 cases of laryngeal carcinoma, but no DNA methylation was found in normal control group, with a significant difference (χ(2) = 15.381, P < 0.05). DNA methylation had no correlation with age, gender, differentiation degree, T stage, pathological type and lymph node metastasis (P > 0.05). The affection of DNA methylation group was more than unmethylation group to expression of gene RASSF1A (t = -3.108, P < 0.01). There was positive correlation between RASSF1A deletion and gene hypermethylation or between H3 lysine 9 methylation of RASSF1A gene promoter and DNA methylation in laryngeal carcinoma tissue(r = 0.816, P < 0.05), but there was negative correlation between H3 lysine 4 methylation of RASSF1A gene promoter and DNA methylation (r = -0.837, P < 0.05) and no correlation between H3 lysine 9 acetylation and DNA methylation (r = -0.383, P > 0.05).

CONCLUSIONSLaryngeal tumor suppressor gene RASSF1A promoter methylation is a key factor down-regulating the gene expression, and histone modifications also plays an important role in tumor development.

Adult ; Aged ; CpG Islands ; DNA Methylation ; Female ; Gene Deletion ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Histones ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm Staging ; Promoter Regions, Genetic ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo investigate the effect of Prostate Water Pellets (PWP) on serum levels of IL-6 and TNF-alpha in rats with chronic bacterial prostatitis (CBP).

METHODSFifty healthy, adult, male Wistar rats with the weight of 180 - 220 g were divided into five groups of ten rats each at random: the control group, model group, high dosage of PWP group, low dosage of PWP group and levofloxacin group. The CBP rat model were created by injecting Escherichia coli (0.2 ml/rat, 10(7)/ml) into prostates. A month later after the model creation, high and low dosage of PWP suspension were used by gavage in CBP rats for 30 days, respectively. Levofloxacin tablets were used by gavage as the positive control, and distilled water was used by gavage in the control and model group. After thirty days, serum levels of IL-6 and TNF-alpha were measured with radioimmunoassay.

RESULTSCompared with the model group, serum levels of IL-6 and TNF-alpha of rats in high and low dosage PWP groups were lower and the difference was significant statistically (P < 0.01).

CONCLUSIONIt has effect to treat CBP rat with the PWP and its mechanism may relate with the decreasing levels of proinflammatory cytokines(IL-6 and TNF-alpha) in blood.

Animals ; Bacterial Infections ; blood ; complications ; drug therapy ; Chronic Disease ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Interleukin-6 ; blood ; Male ; Phytotherapy ; Prostatitis ; blood ; drug therapy ; microbiology ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood

A 20-year-old male patient presented with myalgia of upper limbs and myasthenia of extremities for more than 1 month. Physical examination showed diffuse erythema on the cheeks, upper eyelids, upper chest, neck and dorsa of the hands. The myodynamia of the proximal and distal muscles of upper and lower extremities was grade Ⅳ, Ⅴ, Ⅲ and Ⅴ respectively. Laboratory examinations revealed that the serum levels of creatine kinase, CK-MB and lactate dehydrogenase were 2103 U/L, 83 U/L and 489 U/L respectively, which were all above the normal range. Electromyogram revealed myopathic abnormality and normal nerve conduction velocity. Histopathology of gastrocnemius muscle showed hypertrophy and swelling of muscle fibers, disappearance or fuzziness of transverse striation, and intermuscular lymphoid cell infiltration. A biopsy of the skin lesion from the upper chest showed liquefaction degeneration of and colloid bodies in basal cell layer, perivascular lymphoid cell infiltration in the dermis. A diagnosis of dermatomyositis was established based on the clinical and laboratory findings. After management with intravenous prednisolone 80 mg once daily and symptomatic treatment for 4 weeks, the myodynamia of upper limbs was improved, serum levels of creatine kinase,CK-MB and lactate dehydrogenase reached the normal ranges. However, the myodynamia of lower limbs progressively deteriorated with the emergence of paresthesia. Enhanced MRI scan showed a tumor in the vertebral canal at the level of thoracic vertebra 11 to 12. A spherical encapsulated tumor measuring 3 cm in diameter was surgically removed. The tumor was diagnosed as paraganglioma in vertebral canal according to pathological and immunohistochemical findings. The patient was finally diagnosed with paraganglioma in vertebral canal superimposed on dermatomyositis.

Objective To investigate the effect of microRNA-223-3p (miR-223-3p) on megakaryocytic differentiation and maturation, and explore the potential mechanism .Methods The endogenous expression of miR-223-3p during megakaryocyte ( MK) differentiation was detected by real-time PCR.Flow cytometry further indicated that alteration of miR-223-3p in human cell lines exerted effects on MK differentiation and maturation .By performing integrative bioinformatic analysis, the potential miR-223-3p target gene, MYH10,was identified.Real-time PCR, luciferase reporter assay and flow cytometry revealed that MYH10 was a direct target of miR-223-3p.Results Endogenous expression of miR-223-3p was in-creased with the differentiation and maturation of MK .The expression of megakaryocytic surface markers CD41 and CD61 and the ploidy were significantly increased in K562 and Meg-01 cells after transfection with miR-223-3p mimics.The expression of MYH10 decreased with the increase in miR-223-3p.Using a luciferase reporter assay ,we demonstrated that MYH10 was a direct target of MiR-223-3p.Furthermore, direct downregulation of MYH10 promoted MK polyploidization . Conclusion MiR-223-3p might regulate the polyploidization of MK by targeting MYH10.

The study was aimed to explore the mechanism of SYK and CBL family of ubiquitin ligases in Bufalin-induced HL-60 cells apoptosis. Cell viability was tested by trypan blue staining and apoptosis was detected by using flow cytometry. The expressions of CBL and CBL-b and the phosphorylation of SYK were detected by using immunoprecipitation and Western blot. The results showed that Bufalin inhibited HL-60 cell proliferation in time- and dose-dependent manners. IC(50) of suppressing cell viability at 24, 48 and 72 hours were about 26.3, 7.8 and 2.0 nmol/L respectively. The high dose of bufalin already induced apoptosis of HL-60 cells at 8 hours. SYK was quickly phosphorylated, and the expressions of CBL and CBL-b were down-regulated after treatment with Bufalin. It is concluded that SYK activation and CBL protein down-regulation may be involved in Bufalin-induced HL-60 cell apoptosis.
Apoptosis ; drug effects ; Bufanolides ; pharmacology ; Cell Proliferation ; drug effects ; Down-Regulation ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Proto-Oncogene Proteins c-cbl ; metabolism ; Signal Transduction ; Syk Kinase