Background Glaucoma is the second leading cause of blindness,which is characterized by processing retinal ganglion cells (RGCs) loss and optic nerve dystrophy.Clinical study showed that lowing IOP can not arrest the glaucomatous damage of RGCs.To seek a neuroprotective drug is an urgent need.Objective This present study focused on the effect of triptolide,a natural biologically active compound extracted from Tripterygium wilfordii,on RGCs in glaucomatous eyes. Methods Glaucoma animal models were established in the right eyes of 80 clean Wistar rats by combination with aspiration of aqueous humor and phtocoagulation on anterior chamber angle.Wistar rats were assigned to two groups at random.Triptolide (5μg/kg) was intraperitoneally injected daily from three days before photocoagulation through scarification of animals (total 8 weeks),and same amount of physiologic saline solution was used at the same way.IOP was measured with a Tonopen XL tonometer at at 1day,3,5,7 days and weekly for 8-week duration after phtocoagulation.RGCs numbers was calculated by retinal Nissl staining.Morphology of retina in frozen section was examined under the light microscope.The experiment followed the Standard of Association for Research in Vision and Ophthalmology. Results The IOP was elevated in model eyes from 1 day through 3 weeks after operation with statistically different in comparison with before operation(P<0.05).No obvious differences in IOP changg was found hetween the triptolide group and the normal saline group at each time point(P>0.05).The numbers of RGCs of model eyes in normal saline group decreased gradually after operation,but no evident decline of numbers of RGCs in model eyes in triptolide group. RGCs in triptolide group were considerably more than those of normal saline group in various time points after operation ( P<0. 05). However,no obvious difference in RGCs numbers was found between model eyes and control eyes in Triptolide group. Conclusion Triptolide could protect RGCs in glaucomatous eyes,and its effect does not depend on IOP in chronic glaucoma model.

Objective To investigate the spectrum,diagnosis,time of therapy and management of the congeni-tal heart disease(CHD)in patients with Down′s syndrome(DS).Methods A retrospective report was undertaken of 96 cases in children with DS accompanied by CHD in Department of Pediatric Cardiology,Beijing Anzhen Hospital Af-filiated to Capital Medical University.Data were collected and analyzed about their clinical characteristics,and types of cardiovascular abnormalities,and the important laboratory examinations such as echocardiography and catheterization as well as the procedures of diagnosis and treatments were summarized.Then the interventions,complications and prognosis of different patients were estimated.Results (1)Single congenital heart disease was found in 33 cases (34.38%),a-mong which ventricular septal defect was the most common (14 cases,14.58%),followed by atrioventricular septal de-fect and atrial septal defect (equally,7 cases,7.29%).Multi -cardiovascular abnormalities were discovered in 63 ca-ses,and patent ductus arteriosus turned out to be the most common (42 cases,66.67%).(2)Cat-heterization was car-ried out in 18 cases of serious pulmonary arterial hypertension,and 8 cases were proved resistant pulmonary arterial hy-pertension without operation opportunity.The other 8 cases were estimated as high pulmonary arterial hypertension and medical therapy was suggested before reassessment to reduce surgical risks.(3)Operations were undertaken in 61 ca-ses,among which percutaneous interventional occlusion was performed in 7 cases and surgical interventions were per-formed in 54 patients,in which perioperation complications and death were found in 5 cases and 4 cases,respectively. Conclusions Operation interventions are practicable and most cases recovered well with systematic examinations and assessment in patients with DS and cardiovascular malformations.Early diagnosis and timely interventions are highly suggested.Also close attentions should be paid to follow -up and re -estimation after medical therapy.

Objective To investigate the relationship between serum adiponectin and metabolic syndrome(MS) in children and adolescents with obesity and nonalcoholic fatty liver disease(NAFLD).Methods Four elementary schools and 4 middle schools were selected from Haidian district in Beijing with representative cluster sampling.Two hundred and eighty obese children(obese group),65 obese children with NAFLD(NAFLD group) and 264 normal weight children(healthy control group) aged 7 to 18 years were recruited from the 8 schools with uncompletely randomized sampling.Data including questionnaire,anthropometric measurements,B type ultrasonographic examination for liver were collected and fasting blood laboratory assay were determined.Variables including triglyceride(TG),adiponectin,alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were skewed distribution and natural logarithmical transformations were performed.Chi-square test for category and multiple binary Logistic regression analysis were used to statistical analysis.Results Body mass index(BMI) and waist circumference(WC) in obese group and NAFLD group were higher than those in healthy control group.All the chi-square tests for trend among the 3 groups were statistically significant(P

Cardiomyopathies ; complications ; surgery ; Child ; Female ; Humans ; Male ; Mitral Valve Insufficiency ; etiology ; surgery ; Mitral Valve Prolapse ; Treatment Outcome

To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; drug therapy ; enzymology ; genetics ; physiopathology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Female ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology ; src-Family Kinases ; genetics ; metabolism

Objective: To construct a prediction model for preeclampsia (PE) risk in twin-pregnant women, so as to provide the basis for early screening and prevention of PE. Methods: A total of 467 twin-pregnant women who underwent prenatal examination and delivered at Huzhou Maternal and Child Health Hospital were selected. Sixty cases with preeclampsia (PE) were included in the case group, and 60 women without PE were included in the control group. General information, blood biochemical indicators and uterine artery resistance index (UtA-RI) were collected. A logistic regression model was used to screen predictive factors and establish a nomogram. The Bootstrap method was performed for the internal validation; the receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis were employed to evaluate the discrimination, calibration and clinical utility of the nomogram, respectively. Results: In the case group, there were 47 individuals (78.33%) aged younger than 35 years, 21 individuals (35.00%) with pre-pregnancy body mass index (BMI) of 25 kg/m2 and above, and 33 individuals (55.00%) with in vitro fertilization. In the control group, there were 57 individuals (95.00%) aged younger than 35 years, 8 individuals (13.33%) with pre-pregnancy BMI of 25 kg/m2 and above, and 39 individuals (65.00%) with natural pregnancy. Multivariable logistic regression analysis identified age, pre-pregnancy BMI, method of conception, placental growth factor (PLGF) and UtA-RI as risk prediction factors for PE in twin-pregnant women. The established nomogram had an area under the ROC curve of 0.827 (95%CI: 0.755-0.899), a sensitivity of 0.767, a specificity of 0.733, a good discrimination and calibration, and a relatively high clinical net benefit. Conclusion The nomogram established by age, pre-pregnancy BMI, method of conception, PLGF and UtA-RI has a good predictive value for the risk of PE in twin-pregnant women.

BACKGROUND: Osteoporosis is a genetic disease associated with many enes. To date, the genes that regulate bone mass are incompletely defined.OBJECTIVE: To investigate the relationship between polymorphisms of steoprotegerin (OPG) gene promoter with bone mineral density (BMD) in remenopausal and postmenopausal women.DESIGN: Prospective study.SETTING: Peking Union Medical College Hospital.PARTICIPANTS: In July 2002, 495 Han nationality women selected from Peking Union Medical College Hospital were non-related volunteers and gave their informed consent prior to the study, which included 306 premenopausal women aged 20-39 years, 189 postmenopausal women aged 50-84 years.METHODS: ① BMD measurement: BMD was measured at the Lumbar Spine and Femoral Neck, trochanter, Ward's triangle by dual-energy X-ray absorptiometry. ② Genotyping: Whole blood genome DNA was extracted by QIAGEN DNA extraction kit. The PCR product and the result of endonuclease digest were confirmed by sequencing (Bioasia Biotechnology,Shanghai, China). The impact of the polymorphisms on BMD was also investigated using multiple Logistic regression.MAIN OUTCOME MEASURES: ① Distribution of OPG genotypes and the relationship with BMD. ② Association between OPG polymorphisms and osteoporosis.RESULTS: All 495 subjects were involved in the final analysis. ① These polymorphisms were in Hardy-Weinberg equilibrium (χ2= 0.056 -0.222, P＞ 0.05). The frequencies of genotypes of these subjects were as follows: AA (70.1%), AG (26.9 %), GG (3.0 %) for 163A→G polymorphism; TT (71.3 %), TG (25.9 %), GG (2.8 %) for 245T→G polymorphism. BMD was lower in premenopausal women with GG +AG genotype than AA genotype for 163A→G polymorphism, so did GG+TG genotype than TT genotype for 245T→G polymorphism. But there was no significant difference. BMD was lower in postmenopausal women with AG+GG genotype than AA genotype for 163A→G polymorphism at Lumbar Spine 2-4, Femoral Neck, Ward's triangle and Trochanter (P ＜ 0.05). For 245T→G polymorphism, BMD of postmenopausal women with TG+GG genotype was lower at Femoral Neck,Ward's triangle and Trochanter than TT genotype (P ＜ 0.05). For 245T→G polymorphism, BMD of postmenopausal women with TG+GG genotype was lower at Femoral Neck, Ward's triangle, and Trochanter than TT genotype (P ＜ 0.05). ② Age, weight, height, years since menopause, and 163A→G/245T→G genotypes were sewed as covariates. AG+GG genotype was contributed to low BMD at Lumbar Spine 2-4 and Ward's triangle (OR =2.045, OR=2.956, P ＜ 0.05, 95% CI 1.05-6.7). TG+GG genotype was risk factor for osteoporosis at Lumbar Spine 2-4, Ward's triangle,and Trochanter (OR=2.059, OR=2.859, OR=2.123, P ＜ 0.05, 95% CI 1.04-6.5).CONCLUSION: BMD was lower in postmenopausal women with the variant G allele for 163A→G and 245T→G polymorphisms at Femoral Neck,Ward's triangle, and Trochanter. The variant allele G may associate with lower BMD in postmenopausal women.

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

A gene that could be potentially involved in spermatogenesis was identified and characterized by using suppression subtractive hybridization (SSH) and rapid amplification of cDNA ends (RACE) with total RNA from type A spermatogonia and pachytene spermatocytes of rat. This gene consists of 3 433 base pairs (bp) with a complete open reading frame (ORF) of 3 171 bp and encodes a putative protein containing 1057 amino acids. The nucleotide sequence displays a 93% identity to mouse ubiquitin-activating enzyme E1, Chr Y 1 (Ube1y1) and an 82% identity to human ubiquitin-activating enzyme E1 (UBE1). The putative protein of this gene contains an ubiquitin-activating enzyme signature 1 and an ubiquitin-activating enzyme active site, which are also existed in mouse ubiquitin-activating enzyme E1, human ubiquitin-activating enzyme E1 et al. So we named this gene as Rattus norvegicus ubiquitin-activating enzyme E1 (Ube1). The sequence of Ube1 was submitted to GenBank and the accession number is EF690356. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that Ube1 was specifically expressed in testis, while its expression was not detected in heart, brain, spleen, lung, liver, muscle, kidney and ovary. Comparison of the expression of Ube1 in different developmental stages of testis and Sertoli cells (real-time PCR) indicated that Ube1 was expressed more highly in spermatogonia than in spermatocytes, spermatids and Sertoli cells. In conclusion, Ube1 is a gene encoding rat ubiquitin-activating enzyme E1 and specifically expressed in testis, which might play a key role in ubiquitin system and influence spermatogenesis.
Animals ; DNA, Complementary ; genetics ; Male ; Rats ; Real-Time Polymerase Chain Reaction ; Spermatids ; metabolism ; Spermatocytes ; metabolism ; Spermatogenesis ; genetics ; Spermatogonia ; metabolism ; Testis ; metabolism ; Ubiquitin-Activating Enzymes ; genetics ; metabolism

This study was designed to use iTRAQ technology coupled with 2D LC-MS/MS to study the comparative proteomics of different processing technology for pilose antler. 1015 proteins were identified with 2D LC combined with MOLDI TOF/TOF mass spectrometry. Comparative analysis with Protein Pilot (Version 4.5) revealed that 87 proteins were changed (P ≤ 0.05, the ratio of > 1.50 or < 0.60 as the threshold selection of difference proteins), of which 24 were up regulated and 33 were down regulated in the traditional frying process (TFP) compared with the fresh pilose antler (P ≤ 0.05). 7 significant different proteins (P ≤ 0.001), most of these significantly changed proteins were found to be involved in calcium ion binding and ATP binding associated with human healthy. Freeze drying with protective agent (FDP) (Trehalose) can improve the content of significantly different proteins (P ≤ 0.001) including Collagen alpha-1 (XII) chain (COL12A1) and Collagen alpha-1 (II) chain (COL2A1). The significant function involves in platelets activating, maintenance of spermatogonium, and disorder expression in tumor cells. The functional annotation by Hierarchical clustering and GO (gene ontology) showed that the main molecule functions of the proteins significantly changed in these processes were involved in binding (52.7%), catalytic (25.3%), structural molecule and transporter (6.6%).
Animals ; Antlers ; chemistry ; Chromatography, Liquid ; Collagen ; chemistry ; Down-Regulation ; Freeze Drying ; Gene Expression Regulation ; Proteomics ; Tandem Mass Spectrometry ; Technology, Pharmaceutical ; methods ; Up-Regulation