Animals ; Cell Differentiation ; drug effects ; Cell Transplantation ; Cells, Cultured ; Disease Models, Animal ; Drug Evaluation, Preclinical ; Embryonic Stem Cells ; cytology ; Humans ; Regeneration ; drug effects

OBJECTIVETo study the effect of Tangshenkang Granule (TG) containing serum on renal mesangial cells' (RMCs) proliferation and TGF-β1/Smad2/3 pathway in the high glucose condition.

METHODSTwelve SD rats were randomly divided into four groups, i.e., the low dose TG group, the middle dose TG group, the high dose TG group, and the blank control group, 3 in each group. After 7-day gastrogavage via portal vein blood, rats were sacrificed and their serum samples were collected. RMCs were cultured in common rat serum and TG containing serum respectively. The proliferation of mesangial cells was determined by methly thiazolyl tetrazolium (MTT) assay to determine the optimal TG containing serum concentration. Expression levels of TGF-β1 mRNA and protein were determined by real time quantitative PCR and ELISA. Smad2/3 protein expression and phosphorylation were determined by Western blot and immunofluorescence.

RESULTSTG containing serum at different doses could inhibit high glucose induced RMC cells' proliferation, TGF-β1 over-expression and Smad2/3 phosphorylation.

CONCLUSIONTG containing serum could inhibit high glucose induced RMC cells' proliferation, and its mechanism might be possibly associated with inhibiting TGF-β1/Smad2/3 signaling pathway.

Animals ; Cell Proliferation ; Drugs, Chinese Herbal ; pharmacology ; Glucose ; Mesangial Cells ; Phosphorylation ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Serum ; Signal Transduction ; Smad2 Protein ; metabolism ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo evaluate the antitumor activity of a novel class of 4, 8-Disubstituted-8, 9-dihydropyrazine[2, 3-g]quinazoline-7(6H)-ketones in vitro, and to screen potential anticancer compounds for further study.

METHODSSeventeen compounds of 4, 8-Disubstituted-8, 9-dihydropyrazine[2, 3-g]quinazoline-7(6H)-ketones were synthesized with solid-phase method for biological evaluation of EGFR tyrosine kinase. MTT method was used to evaluate the cytotoxic activity in vitro against three human cancer cell lines (human lung carcinoma cell line A549, human leukemia cell lines K562 and human gastric carcinoma cell line SGC7901).

RESULTSCompound 7-13 and 7-14 showed potent antitumor activities against A549 cells, with IC(50) values of 8.10 and 8.12 mol/L, respectively. Eight compounds showed proliferative inhibition effect on K562 cells, especially 7-2, 7-13 and 7-17, with IC(50) values of 2.22,0.57 and 7.20 mol/L,respectively.And compound 7-13 and 7-3 showed potent antitumor activity against SGC7901 cells, with IC(50) values of 4.20 and 9.71 mol/L, respectively.

CONCLUSIONThe synthesized compounds 4, 8-Disubstituted-8, 9-dihydropyrazine[2, 3-g] quinazoline-7(6H)-ketones show inhibition effects on human cancer cell lines in vitro. Compound 7-13 has anticancer activity in all three cancer cell lines, which might be used as a potential antitumor drug for further study.

Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Humans ; K562 Cells ; Lung Neoplasms ; pathology ; Molecular Structure ; Pyrazines ; chemical synthesis ; chemistry ; pharmacology ; Quinazolines ; chemical synthesis ; chemistry ; pharmacology ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Stomach Neoplasms ; pathology ; Structure-Activity Relationship

AIM:To analyze zebrafish embryos and to identify erythropoietin (Epo) as an active renal anti-apoptotic factor in an Epo receptor (EpoR)-dependent manner. METHODS:For transient knockdown of Epo and EpoR in renal Tg (wt1b:EGFP) zebrafish reporter lines, the morpholino antisense oligonucleotide technique was used. Morphant zebrafish embryos were phenotypically analyzed using fluorescence microscopy. Apoptosis was determined by TUNEL assay and Annexin V staining. Western blot was used to identify Akt phosphorylation in Epo and EpoR knockdown zebrafish. RE-SULTS:Epo and EpoR zebrafish morphants exhibited pathophysiological changes within the pronephros, adversely affecting pronephric structure. Zebrafish embryos upon silencing of Epoa and EpoR showed a significant increase in the apoptosis within the zebrafish pronephros, consequently leading to renal pathophysiological effects. Decreased p-Akt was identified in Epo and EpoR knockdown zebrafish embryos. CONCLUSION:Epo is an essential regulator of renal development and function by interacting with its receptor EpoR and thereby repressing apoptosis, mechanistically by promoting Akt phospho-rylation.

OBJECTIVETo construct a rheumatoid arthritis-specific full-length fully human mammalian display antibody libraries.

METHODSPeripheral blood lymphocytes were isolated from patients with rheumatoid arthritis. The repertoires of kappa light chain (LCκ) and heavy chain variable region (VH) of the antibodies were amplified by RT-PCR. The amplified LCκ and VH genes were inserted into the vector pDGB-HC-TM separately, and the ligated libraries were transformed into competent E.coli TOPO-10 strain to construct the rheumatoid arthritis-specific antibody heavy and light chain libraries. 293T cells were co-transfected with the libraries and the full-length fully human antibody expressed on the surface of 293T cells were analyzed by flow cytometry.

RESULTSThe libraries of rheumatoid arthritis-specific antibody LCκ and heavy chain (IgG1) were constructed. The expression of full-length fully human antibody on the surface of 293T cells was confirmed by flow cytometry. With the rates of correct LCκ and heavy chain sequence insertion reaching 80% and 60%, respectively, as shown by DNA sequence analysis of the randomly selected clones, the libraries showed an expressible combinatory diversity of 6.13×10(10).

CONCLUSIONThe constructed libraries provide a useful platform for screening rheumatoid arthritis-specific antibodies.

Amino Acid Sequence ; Animals ; Antibodies ; genetics ; immunology ; Antibody Specificity ; Arthritis, Rheumatoid ; immunology ; Cell Surface Display Techniques ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; Immunoglobulin Heavy Chains ; biosynthesis ; genetics ; Immunoglobulin kappa-Chains ; biosynthesis ; genetics ; Lymphocytes ; immunology ; metabolism ; Molecular Sequence Data ; Peptide Library ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Transfection

OBJECTIVE: To evaluate the value of early identification and endovascular treatment of iliac vein compression syndrome (IVCS), with or without deep vein thrombosis (DVT). MATERIALS AND METHODS: Three groups of patients, IVCS without DVT (group 1, n = 39), IVCS with fresh thrombosis (group 2, n = 52) and IVCS with non-fresh thrombosis (group 3, n = 34) were detected by Doppler ultrasonography, magnetic resonance venography, computed tomography or venography. The fresh venous thrombosis were treated by aspiration and thrombectomy, whereas the iliac vein compression per se were treated with a self-expandable stent. In cases with fresh thrombus, the inferior vena cava filter was inserted before the thrombosis suction, mechanical thrombus ablation, percutaneous transluminal angioplasty, stenting or transcatheter thrombolysis. RESULTS: Stenting was performed in 111 patients (38 of 39 group 1 patients and 73 of 86 group 2 or 3 patients). The stenting was tried in one of group 1 and in three of group 2 or 3 patients only to fail. The initial patency rates were 95% (group 1), 89% (group 2) and 65% (group 3), respectively and were significantly different (p = 0.001). Further, the six month patency rates were 93% (group 1), 83% (group 2) and 50% (group 3), respectively, and were similarly significantly different (p = 0.001). Both the initial and six month patency rates in the IVCS patients (without thrombosis or with fresh thrombosis), were significantly greater than the patency rates of IVCS patients with non-fresh thrombosis. CONCLUSION: From the cases examined, the study suggests that endovascular treatment of IVCS, with or without thrombosis, is effective.
Adolescent ; Adult ; Aged ; *Angioplasty, Balloon ; *Balloon Dilatation ; Constriction, Pathologic/therapy ; Diagnostic Imaging ; Female ; Humans ; Iliac Vein/*pathology/surgery ; Male ; Middle Aged ; Peripheral Vascular Diseases/complications/diagnosis/*therapy ; Retrospective Studies ; *Stents ; Thrombectomy ; Vascular Patency ; Vena Cava Filters ; Venous Thrombosis/complications/diagnosis/*therapy

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; diagnostic imaging ; pathology ; Female ; Humans ; Lactation ; Male ; Middle Aged ; Ultrasonography

Objective To observe the clinical efficacy of electroacupuncture (EA) at the three groups of Stomach- Meridian acupoints in treating chronic superficial gastritis (CSG) and its effect on serum gastrin level and electrogastrogram, for providing clinical evidence for the experimental result of the three groups of Stomach-Meridian acupoints.Method Sixty-six CSG patients were divided into a treatment group and a control group by using stratified random method, 33 cases in each group. The treatment group was intervened by EA at the three groups of Stomach-Meridian acupoints, while the control group was intervened by EA with ordinary acupoints selection. The intervention was given once a day, successive 6 sessions as a treatment course, for 4 courses in total. The CSG symptoms and syndrome score, serum gastrin content, average amplitude of electrogastrogram, mean waveform frequency, and the percentage of gastric dysrhythmias were observed before and after the intervention, and the clinical efficacies were compared.Result The total effective rate was 90.9％ in the treatment group, versus 87.9％ in the control group, and the between-group difference was statistically insignificant (P>0.05). The total scores dropped significantly in both groups after the treatment (P<0.05), and there was no significant difference in comparing the total score between the two groups after the treatment (P>0.05). The serum gastrin contents declined significantly in both groups after the intervention (P<0.05), and there was no significant between-group difference in comparing the serum gastrin content after the treatment (P>0.05); the average amplitude and mean waveform frequency of preprandial and postprandial electrogastrogram increased significantly in both groups after the treatment (P<0.05), and the percentage of gastric dysrhythmias dropped significantly (P<0.05); after the treatment, there were no significant between-group differences in comparing the average amplitude, mean waveform frequency and the percentage of gastric dysrhy- thmias of preprandial and postprandial electrogastrogram (P>0.05).Conclusion EA at the three groups of Stomach- Meridian acupoints can effectively improve the symptoms, down-regulate serum gastrin, help gastric motility, and restore gastric rhythm of CSG patients; its efficacy is equivalent to that of the classic acupoints group, i.e. Neiguan (PC6), Zhongwan (CV 12), and Zusanli (ST 36).

OBJECTIVETo investigate the expression of Junctophilin 1 (JP1) in cardiogenesis of mammalian.

METHODSCardiac differentiation of embryonic stem cells (ESCs) was generated by hanging drop method. Fetal heart was obtained from the rats aged d 14-20 of gestation. The expression of JP1 and JP2 during cardiogenesis of ESCs and rat embryos was analyzed by RT-PCR or Western blotting. Immunofluorescence staining was employed to reveal the distribution of JP1 and JP2 in embryoid body (EB), probing for merging of JP1 and JP2 and cardiac sarcomeric α-Actinin or Troponin-T. Percentage of JP1 and JP2-positive staining cells was analyzed quantitatively by FCS on d17.

RESULTSJP1 mRNA was up-regulated at the early stage (d 5-11) and then decreased. The expression of JP1 protein was up-regulated at the early stage (d 7-9), then decreased gradually and disappeared after d 15. While JP2 gene and protein expression increased in a time-dependent manner during cardiogenesis of rat embryos. The results of immunofluorescence staining showed that there was a parallel co-localization of JP2 with Troponin-T or α-Actinin on d17, while JP1 failed to express in the sarcomeric positive area at the same time point. Furthermore, FCS analysis showed that about 16.59% of cells were JP2-positive, while no cells were stained positively for JP1 in d17 EBs.

CONCLUSIONJP1 gene is expressed during the whole process of cardiogenesis, while JP1 protein only appears on the early stage. The expression of JP1 in cardiogenesis of ESCs is consistent with that of rat embryos.

Actinin ; genetics ; metabolism ; Animals ; Cell Differentiation ; Cell Line ; Embryonic Stem Cells ; cytology ; metabolism ; Heart ; embryology ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Inbred ICR ; Myocytes, Cardiac ; cytology ; metabolism ; RNA, Messenger ; genetics ; Rats ; Troponin T ; genetics ; metabolism

Objective To understand the status of health literacy of residents in Qiandao Lake town in order to complete the surveillance system.Methods A total of 484 residents aged 15 to 69 years old from 9 communities were selected using cluster random sampling method.Then the household interview was conducted.Results The overall level of health literacy of residents was 26.03%.As for the 4 health literacy related aspects including basic health knowledge,health lifestyle and behaviors,health related skill and health beliefs,the rates were 32.54% ,4.77% ,60.30% and 92.62%,respectively. The multivariate logistic regression analysis showed that the level of health literacy of residents was related to age and education.Conclusion The health education in Qiandao Lake town has got some achievements.