Objective To compare the effects of vitrification with slow-freezing on the developmental ability of day 3 cleavage stage embryos.Methods Patients who had no less than 4 high quality embryos were included in this study.These embryos were cryopreserved using the methods of vitrification or slow-freezing.In the eryopreserved embryo transfer cycles,the embryos which were cryopreserved using one of the methods were chosen randomly.The developmental ability of embryos was compared between these two groups.Results A total of 80 patients were included in this study with 160 embryos.In the group of slow-freezing,73(91%)embryos were survived and achieved 15(38%)clinical pregnancies.Among these,3 were twins and the implantation rate was 25%(18/73).In the group of vitrification,71(89%)embryos were survived and achieved 19(48%)clinical pregnancies.Among these, 9 were twins and the implantation rate was 39%(28/71),which was significantly higher than the slow- freezing group(P

Background Retinitis pigmentosa (RP) is a monogenic inheritance and blinding disease of fundus oculi.There is not an effective therapeutic method now.Objective This work was to identify the mutations of RP1 gene in Chinese RP patients in Ningxia area and to explore the potential interactions in the pathogenesis of RP.Methods The periphery blood of 3-5 ml was collected from 110 individuals with RP(35 ADRP and 75SRP)and 100 normal controls in Ningxia area.Polymerase chain reaction (PCR) and direct DNA sequencing were used to screening the sequence alterations in the entire coding region and splice sites of RP1 gene.Multivariate analysis and two web-based programs( PolyPhen and SIFT) were used to analyze the results.Results Eleven mutation locus were detected in the exon 4 of RP1 gene including two novel sequence variants:p.Lys1152Lys without a higher mutation rate in comparison with normal control group(x2 =9.12 P＜0.01 ),but c.* 247A＞C with a higher mutation rate in comparison with normal control group(x2 =12.77,P＜0.01 ) and c.* 247A＞C mutation was thought to be correlated with RP( r=1.11,P＜0.05 ).The other ten mutation locus were reported as single nucleotide polymorphisms (SNP).The mutation rate of p.Gln1725Gln was found to be higher in the RP patients than the normal controls (x2 =42.09,P＜0.01 ),but no the significant correlation was seen between the pathogenesis of RP and mutation of p.Gln1725Gln(r=1.74,P＞0.05).p.Lys1152Lys mutation was found in only 1 patient.Three SNPs( p.Arg872His,Ala1670Thr,Ser1691Pro) were always occurred in the same 83 RP patient and the relevance ratio was higher than controls ( P＜0.01 ).The age of night blindness on patients with concurrent three mutations was (30.54± 13.68 ) years,and the best corrected visual acuity (BCVA) was 0.50 ± 0.38.The age of night blindness on patients without concurrent three mutations was(21.06± 16.24) years,and the BCVA was 0.40 ±0.33 and were higher than controls ( t =2.11,P ＜ 0.05 ).Conclusions In this study,the prevalence of RP1 mutations among the RP patients in Ningxia population was lower than other populations (＜ 1％ ).The alliance of SNPs (p.Arg872His、p.Ala1670Thr、p.Ser1691Pro) may play a protective role on RP patients and reduce the frequency of mutatiaon in RP1 gene.

OBJECTIVETo investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men.

METHODSWe collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease.

RESULTSQuality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group.

CONCLUSIONCalcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

Calcimycin ; therapeutic use ; Calcium Ionophores ; therapeutic use ; Female ; Humans ; Infertility, Male ; drug therapy ; Male ; Oocytes ; Pregnancy ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; abnormalities

Objective To determine whether the susceptibility to ischemia-reperfusion injury in aged heart is higher than that in adult heart and,if so,to clarify the mechanisms underlying this change.Methods Wister rats(5-or 20-month-old)were randomly divided into 4 groups(6 animals in each group).The rats were subjected to 30 minutes of myocardial ischemia via ligating the left anterior descending coronary artery,followed by 3 hours of reperfusion(Young-MI/R group and Old-MI/R group);A silk suture around the left anterior descending coronary artery was not ligated in young and old rats(Young-sham group and Old-sham group).Myocardial apoptosis was detected by terminal deoxynueleotidyl transferase biotin-d UTP nick end labeling(TUNEL)staining and caspase-3 activity was detected by using a caspase-3 colorimetrie assay.Nitrotyrosine content,a footprint of in vivo ONOO~-formation,and total NO content were determined by ELISA and chemiluminescence method respectively.Results A significantly exacerbated cardiac reperfusion injury was found in Old-MI/R group as evidenced by increased TUNEL positive myocytes[(19.0?2.1)% vs.(14.6?1.7)%],and increased myocardial caspase-3 activity[(436?35)?mool/mg vs.(340?32)?mol/mg] compared with Young-MI/R group(P＜0.05).Aged hearts had a markedly increased basal NOx level compared with young adult hearts.Marked higher myocardial nitrotyrosine content was found in OId-MI/R group[(7.25?0.18)nmol/g]than that in Young-MI/R group[(4.68?0.15)nmol/g] (P＜0.05).Conclusions In aged hearts,high levels of NO might form highly toxie derivant, ONOO~-,and its subsequent nitrified protein.This may attribute to the increased susceptibility of the aged heart to isehemic-reperfusion injury.

Based on data related to human brucellosis which was collected from the national notifiable infectious disease reporting system in the 6 provinces(Inner Mongolia,Shanxi,Heilongjiang,Shaanxi,Jilin and Liaoning)of north China from 2004 to 2007,at the county scale.Data would include age and gender standardized mortality ratios(SMRs)while ESDA was including histograms,box plots and box maps,global and local Moran' s I statistics,etc.The global Moran' s I values from 2004 to 2007 were 0.2581,0.4574,0.4457,0.4841,respectively and all with statistically significant differences.Most of local Moran' s I values were significant positive statistically.High-high counties were mainly in the northeast,most of which were pastoral areas,but the farming-pastoral areas and agricultural areas/town had an increasing trend over time.Low-low counties were mainly in the western and southern areas and most of which were agricultural areas/towns.Low-high counties appeared to be rare,mainly around the counties with high incidence,mainly belonged to agricultural areas/towns.The incidence rates of brucellosis in the six provinces of north China had a trend of increase from 2004 to 2007,namely spreading from east to west,from south to north,and from pastoral areas to farming-pastoral areas and agricultural areas/towns.ESDA could be used to develop effective measures for prevention and control of brucellosis.

Objective To explore the relationship between the polymorphisms of exon 41 in COL6A3 and sporadic isolated dystonia in China. Methods A total of 127 outpatients with isolated dystonia and other 130 age-and gender-matched healthy controls were collected their blood samples. The single nucleotide polymorphism (SNP) was screened from 1000 Genomes Project. Genotype was detected with polymerase chain reaction-restriction fragment length polymorphism and the genotype and allele distribution were compared between the patients and the controls. Results Two SNPs in exon 41 in COL6A3 were found,named rs1131296 and rs2270669.There was no difference be-tween the patients and the controls in both genotype and allele(χ2<1.829,P>0.05).There was no difference in the age of onset among the patients with various genotypes(P>0.05). Conclusion Polymorphism of exon 41 in COL6A3 gene may not contribute to risk of sporadic isolated dystonia in China.

OBJECTIVETo investigate the effect of baicalein on proliferation and migration of multiple myeloma (MM) cell lines and its molecular mechanism.

METHODSThe MM cell line RPMI-8226 and U266 cells were used as the model, and treated with different concentration and time of baicalein the effect of baicalein on the MM cells proliferation was assessed by MTT assay. With or without baicalein or Interleukin-6 (IL-6) treatment, the β-catenin protein level was analyzed by immunofluorescence assay and western blot assay and mRNA levels of β-catenin, c-myc, cyclin D1 and integrin 7 gene by RT-PCR. Transwell chamber migration assay was used to detect the cells migration ability with different concentration of baicalein cultured.

RESULTSBaicalein inhibited the MM cell line RPMI 8226 and U266 cell proliferation in a dose- and time-dependent manner. It simultaneously inhibited β-catenin protein level to resist the effect of IL-6 on inducing MM cell proliferation, and resulted in decrease of β-catenin, c-myc, cyclinD1 and integrin β7 mRNA levels. Baicalein also decreased migration ability of MM cells in a dose-dependent manner by SDF-1.

CONCLUSIONBaicalein can inhibit MM cells proliferation and migration, and its molecular mechanisms are associated with inhibition of proliferation related genes β-catenin, c-myc, cyclin D1 and integrin β7 expression.

Antigens, CD ; metabolism ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Flavanones ; pharmacology ; Humans ; Integrin alpha Chains ; metabolism ; Interleukin-6 ; pharmacology ; Multiple Myeloma ; metabolism ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; RNA, Messenger ; genetics ; beta Catenin ; metabolism

BACKGROUNDGalectin-3 is the most recently identified advanced glycosylation end products (AGEs) binding protein. This study aimed to investigate the effects of AGEs and rosiglitazone on the expression and secretion of galectin-3 in cultured human renal mesangial cells (HRMCs).

METHODSHRMCs were incubated with different concentrations of AGE-bovine serum albumin (BSA) (0, 50, 100, 200, and 400 mg/L) for different time (0, 24, 36, 48, and 72 hours), and exposed to AGE-BSA in the presence of different concentrations of rosiglitazone (1, 10, and 100 micromol/L). The mRNA and protein expression of galectin-3 in HRMCs were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. The culture medium of HRMCs was collected and concentrated, and the content of galectin-3 in the medium was detected by Western blotting.

RESULTSBoth RT-PCR and Western blotting revealed that AGE-BSA up-regulated the expression of galectin-3 in HRMCs in a concentration- (P < 0.05) and time-dependent (P < 0.05) manner compared with the control. Compared with the control, AGE-BSA elevated the content of galectin-3 in the culture medium of HRMCs time- and concentration-dependently (P < 0.05, respectively). Both protein and mRNA expression of galectin-3, and its content in the medium of HRMCs exposed to different concentrations of rosiglitazone in the presence of AGE-BSA were increased compared with those of cells exposed to AGE-BSA alone (P < 0.05). Rosiglitazone increased the expression and secretion of galectin-3 in a dose-dependent manner (P < 0.05).

CONCLUSIONSAGEs up-regulates the expression and secretion of galectin-3 in HRMCs. Rosiglitazone further enhances the upregulation of galectin-3 in HRMCs induced by AGEs, which suggests that rosiglitazone may play a role of reno-protection via up-regulation of galectin-3.

Blotting, Western ; Cell Line ; Galectin 3 ; genetics ; secretion ; Glycation End Products, Advanced ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Mesangial Cells ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Serum Albumin, Bovine ; pharmacology ; Thiazolidinediones ; pharmacology

OBJECTIVETo investigate the effect of compound qingqin liquid (CQL) on Toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in rats with urate nephropathy, and to explore its renal protection mechanism.

METHODSTotally 55 SD rats were randomly divided into 5 groups, i.e., the normal control group (n =5), the model group (n =10), the positive drug group (n=10), and the high-, medium-, low-dose CQL groups (n=10) respectively. The urate nephropathy model was induced by intragastrically administering adenine and feeding yeast. Distilled water was intragastrically administered at the daily dose of 10 mL/kg to rats in the normal control group and the model group. Allopurinol was intragastrically administered at the daily dose of 9.33 mg/kg to rats in the positive control group. CQL was intragastrically administered at the daily dose of 3.77, 1.89, 0.94 g/kg to rats in the high-, medium-, and low-dose CQL groups. Rats of each group were executed in batches at the 4th and 6th week respectively. Their kidney tissues were taken out to determine the mRNA transcription level of TLR2 and TLR4 by reverse transcription-polymerase chain reaction (RT-PCR). The protein expression level of TLR2 and TLR4 were determined by Western blot. The protein expression level of TLR4 was also detected by immunohistochemical assay.

RESULTSAt week 4 and 6, the protein expression of TLR2 and TLR4 as well as the mRNA transcription of TLR4 increased in the model group, when compared with the control group (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in the transcription level of TLR2 mRNA or TLR4 mRNA among the 3 CQL groups (P > 0.05) at week 4 and 6. Additionally, at week 6, the protein expression of TLR4 and TLR2 could be reduced by CQL (P < 0.05, P < 0.01).

CONCLUSIONCQL might protect kidney tissue against inflammatory injury by inhibiting the protein expression levels of TLR2 and TLR4.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Kidney ; drug effects ; metabolism ; Kidney Diseases ; drug therapy ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; Uric Acid

Twenty-two compounds were isolated from the flowers of Scabiosa tschilliensis. Their structures were identified by spectroscopic methods as octacosanol (1), stearic acid (2), β-sitosterol (3), oleanolic acid (4), apigenin (5), luteolin (6), daucosterol (7), kaempferol-3-O-β-D-6-O-(p-hydroxycinnamoyl) -glucopyranoside (8), kaempferol-3-O-β-D- (3, 6-di-p-(hydroxycinnamoyl) -glucopyranoside (9), apigenin-7-O-β-D-glucopyranoside (10), luteolin-4'-O-β-D-glucopyranoside (11), apigenin-7-O-rutinoside (12), luteolin-7-O-β-D-glucopyranoside (13), apigenin-4'-O-β-D-glucopyranoside (14), caffeic acid methyl ester (15), loganin (16), adenosine (17), luteolin-6-C-β-D-glycopyranosyl (18), sweroside (19), sylvestrosides I (20), sylvestrosides II (21), urceolide (22). Among them, compounds 1, 2, 7-9, 12, 15, 17-18, 20-22 were isolated from the genus Scabiosa for the first time, and compounds 1-4, 6-9, 11-12, 14-22 were isolated from this plant for the first time. 13C-NMR data of 22 were reported for the first time.
Dipsacaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Flowers ; chemistry ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization