Animals ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Endothelial Cells ; cytology ; drug effects ; physiology ; Female ; Humans ; Mice ; Mice, Inbred C57BL ; Neovascularization, Physiologic ; drug effects ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVEThe aim of this study was to investigate the effect of 5-aza-2'-deoxycytidine (5-Aza-dc), a methylation inhibitor, on cisplatin-resistance in non-small cell lung cancer cell line A549/DDP and to explore its possible mechanism.

METHODSMTT assay was used to test the cytotoxicity of 5-Aza-dc on A549/DDP cells, and the IC(50) and cisplatin resistance index of A540/DDP cells at 48 hours after 5-Aza-dc (0 µmol/L, 20 µmol/L, 40 µmol/L) treatment at different concentrations. MSP, fluorescence quantitative RT-PCR (real-time RT-PCR) and Western blot were used to detect the hMLH1 methylation status, mRNA and protein expressions, respectively.

RESULTSThe IC(50) value of cisplatin in A549/DDP cells was 30.15 ± 0.76 µmol/L. The MTT assay results demonstrated that during the 5-Aza-dc treatment for 48 hours, the dose of 20 µmol/L was non-toxic and 40 µmol/L was low-toxic. 5-Aza-dc at those two doses reduced IC(50) value of cisplatin to 16.54 ± 0.35 µmol/L (RI = 1.82) and 6.82 ± 0.16 µmol/L (RI = 4.42), respectively. MSP, real-time RT-PCR and Western blot showed that 5-Aza-dc at non-toxic and low-toxic doses removed the partial hMLH1-hypermethylation, and up-regulated hMLH1 mRNA and protein expressions.

CONCLUSIONSLow dose 5-Aza-dc can partially reverse the cisplatin-resistance in A549/DDP cells, which may be achieved through removal of hMLH1 hypermethylation and increased expression of hMLH1 gene. 5-Aza-dc may have a role in increasing the efficacy of chemotherapy for patients whose tumors are lack of hMLH1 expression because of hMLH1 promoter hypermethylation.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Antimetabolites, Antineoplastic ; administration & dosage ; pharmacology ; Azacitidine ; administration & dosage ; analogs & derivatives ; pharmacology ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; DNA Methylation ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; MutL Protein Homolog 1 ; Nuclear Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; metabolism

In order to reaction the quality present situation, problems on the current quality of animal sources of drugs are summed up by using test data analysis, literature search and marketing research. This paper can also help the improvement of the quality management, the revision of the relevant department policy system and the improvement of standards.
Animals ; China ; Medicine, Chinese Traditional ; standards ; Pharmaceutical Preparations ; analysis ; standards ; Quality Control

Objective To study the clinical significance of the serum angiopoietin-2(Ang-2) in the diagnosis,recurrence,invasion and metastasis of gastric cancer.Methods The serum Ang-2 and carcino-embryonic antigen (CEA) levels in 158 patients with gastric cancer (gastric cancer group) and 30 normal controls(control group) were measured by enzyme linked immunosorbent assay(ELISA) technique respectively.The serum Ang-2 and CEA levels were also measured 2 weeks after operation in gastric cancer group and reexamined in the recurred gastric cancer patients in 2 years after operation (recurred and metastasis group).The correlation between the serum Ang-2 level and pathologic c haracterization of gastric cancer was evaluated.Results The serum Ang-2 and CEA levels in gastric cancer group [ (331.8 ±64.3),(42.6 ±37.3)μg/L] and recurred and metastasis group [(318.7 ±72.9),(40.5 ±36.7)μg/L] were significantly higher than those in control group [ (187.4 ± 32.7),(4.2 ± 3.1 )μ g/L] (P ＜ 0.01 ),and the serum Ang-2 level 2 weeks after operation [ (211.6 ± 75.1 ) μ g/L ] was significantly decreased to the control group (P ＞ 0.05 ),while the serum CEA level [ (33.4 ± 30.6) μ g/L ] was still significantly higher than the control group (P ＜ 0.01 ).The sensitivity of the serum Ang-2 for diagnosis of gastric cancer was markedly higher than that of the serum CEA (P ＜ 0.01 ).There was correlation between serum Ang-2 and degree of tumor differentiation,TNM pathological staging,lymphatic metastasis,invasion depth and tumor size (p ＜0.01 ),but there was no correlation between serum Ang-2 and tissue classification and location of gastric cancer (P＞ 0.05).Conclusion The serum Ang-2 level is suggested to be a valuable gastric cancer marker and conduce to the diagnosis of gastric cancer,the monitoring of recurrence after operation and evaluation of prognosis.

Objective: To observe the effect of electroacupuncture (EA) pretreatment on the protein expression of c-fos in fastigial nucleus (FN) and lateral hypothalamus area (LHA) in rats with acute myocardial ischemia-reperfusion injury (MIRI), and to explore the role and mechanism of FN and LHA in EA at the Heart Meridian fighting against acute MIRI reaction. Methods: Seventy Sprague-Dawley rats were randomly divided into a sham operation group, a model group, an EA-Heart Meridian group and an EA-Lung Meridian group, with 14 rats in each group; an LHA lesion plus EA-Heart Meridian group (LHA+EA-Heart Meridian group) and a FN lesion plus EA-Heart Meridian group (FN+EA-Heart Meridian group), with 7 rats in each group. Except the sham operation group, the left anterior descending branch of coronary artery was ligated to establish acute MIRI rat models in the other 5 groups. In the three groups with EA-Heart Meridian treatment, Shenmen (HT 7) and Tongli (HT 5) were selected; Taiyuan (LU 9) and Lieque (LU 7) were selected in the EA-Lung Meridian group. All the EA groups received EA stimulation prior to modeling, with 1 mA in current intensity and 2 Hz in frequency, 20 min each time, once a day for a total of 7 d. The sham operation group and the model group did not receive EA stimulation. The electrocardiogram was observed in the rats to analyze the ST-segment deviation and cardiac arrhythmia score. The expression of c-fos protein in FN and LHA was detected by immunohistochemistry method. Results: Compared with the sham operation group, the ST-segment deviation, cardiac arrhythmia score and the expression of c-fos protein in the FN and LHA increased significantly in the model group (all P<0.05). Compared with the model group, the ST-segment deviation, cardiac arrhythmia score and the expression of c-fos protein in FN and LHA decreased significantly in the EA-Heart Meridian group (all P<0.05). Compared with the EA-Heart Meridian group, the ST-segment deviation and cardiac arrhythmia score increased significantly in the EA-Lung Meridian group, LHA+EA-Heart Meridian group and FN+EA-Heart Meridian group (all P<0.05); the expression of c-fos in FN increased significantly in the EA-Lung Meridian group and LHA+EA-Heart Meridian group (both P<0.05); the expression of c-fos in LHA increased significantly in the EA-Lung Meridian group and FN+EA-Heart Meridian group (both P<0.05). Conclusion: FN and LHA are involved in the mechanism of EA at Heart Meridian to improve the acute MIRI reactions, and the cerebellum may participate in the improvement of cardiac function by EA through the cerebellum-hypothalamus projection.

OBJECTIVETo investigate the frequency and type of PHEX gene mutations in children with X-linked hypophosphatemic rickets (XLH), the possible presence of mutational hot spots, and the relationship between genotype and clinical phenotype.

METHODSClinical data of 10 children with XLH was retrospectively reviewed. The relationship between gene mutation type and severity of XLH was evaluated.

RESULTSPHEX gene mutations were detected in all 10 children with XLH, including 6 cases of missense mutation, 2 cases of splice site mutation, 1 case of frameshift mutation, and 1 case of nonsense mutation. Two new mutations, c.2048T>C and IVS14+1delAG, were found. The type of PHEX gene mutation was not associated with the degree of short stature and leg deformity (P=0.571 and 0.467), and the mutation site was also not associated with the degree of short stature and leg deformity (P=0.400 and 1.000).

CONCLUSIONSMissense mutation is the most common type of PHEX gene mutation in children with XLH, and c.2048T>C and IVS14+1delAG are two new PHEX gene mutations. The type and site of PHEX gene mutation are not associated with the severity of XLH.

Adolescent ; Child ; Child, Preschool ; Familial Hypophosphatemic Rickets ; genetics ; Female ; Humans ; Infant ; Male ; Mutation ; PHEX Phosphate Regulating Neutral Endopeptidase ; genetics ; Retrospective Studies

The fruit shape is important quantitative trait closely related to the fruit quality. However, the genetic model of fruit shapes has not been proposed. Therefore, in the present study, analysis of genetic effects for fruit shape traits (fruit length and fruit perimeter) in sponge gourd was conducted by employing a developmental genetic model including fruit direct effects and maternal effects. Analysis approaches of unconditional and conditional variances were applied to evaluate the genetic behavior of fruit shape traits at economical and physiological maturation times. The results of variance analysis indicated that fruit length and fruit perimeter were simultaneously affected by fruit direct genetic effects and maternal effects. Fruit direct genetic effects were relatively more important for fruit shape traits at whole developmental period. The gene expression was most active at the economical maturation stage (1 approximately 12 d after flowering) for two shape traits, and the activation of gene was mostly due to direct dominance effects at physiological maturation stage (13 approximately 60 d after flowering). The coefficients due to different genetic effects, as well as the phenotypic correlation coefficients, varied significantly between fruit shape traits themselves at various maturation stages. The results showed that it was relatively easy to improve fruit shape traits for industrial purpose by carefully selecting the parents at economical maturation stage instead of that at physiological maturation stage.
Computer Simulation ; Fruit ; anatomy & histology ; physiology ; Luffa ; anatomy & histology ; physiology ; Models, Genetic ; Quantitative Trait, Heritable

Objective To approach the clinical significance of Clara cell secretary protein(CCSP) in bronchial asthmatic children. Methods Serum were collected from 50 cases during asthmatic attacks, 22 asthmatic children who were in stable conditions, and 20 healthy children. Serum CCSP concentrations were measured by a human CCSP enzyme- linked immunosorbent assay (ELISA). Results Asthmatic children had significantly lower levels of CCSP in serum during asthmatic attacks(P

Objective To explore the role of Clara cell secretory protein(CCSP) in asthmatic children and compare the levels of CCSP in sera and induced sputum.Methods Thirty-four children with asthma who were in remission and 25 healthy controls were enrolled.Sera and hypertonic saline-induced sputum were obtained in asthmatic children,and sera alone were obtained in control subjects.The le-(vels) of CCSP were measured in sera and induced sputum by enzyme linked immunosorbent assay.Results Asthmatic children,compared with controls,had significantly lower concentration of CCSP in sera(P

0.05).In addition,in different medication intervals and the same total dosage(200 mg),there was no difference in the number of patients who reached ASAS20,ASAS50 anti BASDAI50 in both groups.The changes of other parameters were not observed.Conclusion Two dosages and different medication interval of rhTNFR-Fc have similiar efficacy onset time and maintenee period.Mean- while,at the same total dosage,there is no signifieant difference in therapeutic effect in the two dosage groups. However,50 mg(1/7 d)regimen has better compliance than 25 mg(1/3 d).