Diosgenin can inhibit the growth of A375 and K562 cell lines and induce their apoptosis with an effect on pro-apoptotic members of Bcl-2 family. To study the SAR of diosgenin derivatives, and to improve the anti-tumor activity of diosgenin, a series of novel diosgenin derivatives were designed and synthesized. Their anti-tumor activities in vitro were evaluated. The results revealed that most of the new derivatives had potent effects against K562, A375 and A549 (three tumor cell lines) in vitro, and had no or less effect against H293 and L02 (two normal cell lines). Particularly, some compounds (e.g. 1, 6-8) showed excellent activities on K562 with IC50 values ranging from 1.96 to 4.35 micromol x L(-1).
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Diosgenin ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Drug Design ; Humans

Objective: To evaluate the associative role of depression and apolipoprotein E epsilon 4 allele (APOEε4) in subjective cognitive decline (SCD) and its progression to objective cognitive decline. Methods: After literature search in electronic databases, studies were selected by following precise eligibility criteria. Meta-analyses were performed to examine the role of APOEε4 and depression in SCD or its progression to mild cognitive impairment (MCI) or dementia. Results: APOEε4 positivity was not different between SCD and normal individuals but was significantly higher in individuals with SCD plus than in normal individuals [odds ratio: 2.39 (95% CI: 1.87, 3.05); p<0.00001] and in SCD converters than in non-converters [odds ratio: 5.19 (95% CI: 2.36, 11.42); p<0.00001]. Depression was significantly higher in individuals with SCD [standardized mean difference: 0.63 (0.45, 0.82); p<0.00001] and SCD plus [standardized mean difference: 0.83 (0.43, 1.22); p<0.0001] than in normal individuals. However, depression was not different between SCD and MCI or between SCD converters and non-converters. Age of SCD converters was higher than non-converters [mean difference: 2.95 years (0.58, 5.31)]. Conclusion Whereas APOEε4 positivity was higher in SCD plus and SCD converters, depression was higher in SCD and SCD plus but was not different between SCD and MCI.

OBJECTIVETo construct the Gcn5 shRNA plasmids and to explore the Gcn5 shRNA role in histone acetylation modification with the differentiation of stem cells.

METHODSSeven shRNA fragments were recombined into pGenesil-1 vector to form 7. Gcn5 shRNA constructions. The mesenchymal stem cells (MSCs) induced for two weeks with 5-aza were transfected by the plasmids with lipofectamine2000. Polyclonal antibodies labeled with TRITC were used to identify the acetylation in MSCs with or without Gcn5 shRNA constructions. The efficiencies of transfection and RNAi were calculated based on the ratio of GFP (green fluorescence)/DAPI (blue fluorescence) and TRITC (red fluorescence)/DAPI, respectively.

RESULTSSeven Gcn5 shRNA plasmids or constructions were identified by restriction endonucleases Pst I/Sal I and DNA sequencing. Acetylation block was observed after Gcn5 shRNA plasmids transfected into cells. Fluorescent intensity of TRITC in nucleuses were decreased remarkably, or even disappeared in MSCs. The efficiencies of transfection and RNAi were 93.7% and 46.6%, respectively.

CONCLUSIONThe Gcn5 shRNA plasmids constructed in the present study can decrease the histone acetylation during cell differentiation. It sets the basis for further exploring the role of acetylation in the regulation of cell differentiation.

Acetylation ; Animals ; Cell Cycle Proteins ; genetics ; Cell Differentiation ; drug effects ; physiology ; Histone Acetyltransferases ; genetics ; Histones ; metabolism ; Plasmids ; genetics ; RNA Interference ; RNA, Small Interfering ; pharmacology ; Rats ; Rats, Wistar ; Stem Cells ; cytology ; drug effects ; Transcription Factors ; genetics ; p300-CBP Transcription Factors

This study was carried out to examine the effect of different donor cell type and micro-manipulation on the development of reconstituted embryos. Cultured mural cumulus cells or fibroblast cells from an adult transgenic goat expressing human erythropoietin(rhEPO) were used as the donor cells in nuclear transfer experiments. The reconstituted eggs were generated by transferring fibroblast cells or cumulus cells into the perivitelline space of enucleated M II oocytes and then followed by electrofusion and activation. After 6 days' incubation in vivo, the reconstructed embryos developed into morulae or blastocysts were transferred into 6 foster recipients. Two of the foster-mothers were pregnant and gave birth to two offspring, which were derived from the fibroblast cell and cumulus cell, respectively. Fingerprint analysis showed that the PCR-RFLP patterns of the two offspring were identical to that of donor goats. PCR results indicated that these cloned goats carried hEPO gene as same as their donor cells.
Animals ; Animals, Genetically Modified ; genetics ; Cell Fusion ; methods ; Cloning, Organism ; Embryo Transfer ; trends ; Erythropoietin ; biosynthesis ; genetics ; Fibroblasts ; cytology ; Goats ; embryology ; genetics ; Humans ; Microinjections ; methods ; Nuclear Transfer Techniques ; Oocytes ; cytology

OBJECTIVETo retrospectively analysis the curative effect of wrist scaphoid bone fracture,and explore the causes and preventive methods of misdiagnosis.

METHODSFrom September 2007 to September 2010,16 patients with wrist scaphoid bone fractures were treated with plaster cast and cannulated screws fixation. There were 10 males and 6 females,ranging in age from 26 to 44 years with an average of 35 years. Among them, 12 cases manifested swelling pain of radial lateral wrist, tenderness at snuffbox area, wrist pain aggravated when stretching wrist joint, thumb or forefinger; 4 cases manifested no obviously symptoms and limited movement; 9 cases were early diagnosed; 5 cases were treated by plaster cast; 4 cases were treated with cannulated screws fixation; Among 7 cases with misdiagnosis, there were 4 cases without obvious symptoms and they were dealt with activating blood to dissipate swelling and pain process in preliminary stage. Four cases were treated with plaster cast and 3 cases with cannulated screws fixation.

RESULTSAll the patients were followed up from 3 months to 39 months (averaged 21 months). Among 16 patients, 9 cases were early diagnosis, 7 cases were misdiagnosis and the rate of misdiagnosis was 43.8%. Seven cases with screws fixation were no wound infection. There was 1 case with occurred chronic pain and declining wrist mobility in both plaster cast and screw group, and both of them were misdiagnosed. According to curative effect rating criteria,these 2 cases were classified into moderate, other 14 cases were excellent.

CONCLUSIONWrist scaphoid bone fracture are easy to misdiagnose, so early diagnosis and treatment is particularly important. The main causes of misdiagnosis are nonspecific symptoms at early stage, combination with other injuries, lack of knowledge and ignorance of the further examination. Therefore, detailed inquiries and particular examination, multi-dimensional radiography and CT scan or MRI scan are the main measures for prevention.

Adult ; Bone Screws ; Casts, Surgical ; Diagnostic Errors ; prevention & control ; Female ; Fractures, Bone ; diagnosis ; surgery ; Humans ; Male ; Retrospective Studies ; Scaphoid Bone ; injuries

Objective To investigate the molecular characteristics of Japanese encephalitis virus (JEV) living in vector mosquitoes,from Zhejiang province.Methods A total of 13620 mosquitoes were collected from the monitoring stations located in Cixi city and Xianju county in Zhejiang province,in July and August,2009-2010.Nucleic acid of JEV from the mosquitoes was monitored by using real-time RT-PCR.The virus strains were isolated with BHK-21 cell line,with E genes of the isolated viruses amplified,sequenced and their phylogeny and homology analyzed.Results The positive rates of JEV for those mosquitoes collected in the stations of Cixi and Xianju were 17.0％ (27/159) and 3.4％ ( 1/29 ),respectively.Twenty-two JEV strains were isolated,accounted for 15.4％ among the 143 batches of mosquitoes collected in 2010.All E genes in the 6 sequenced virus isolates contained 1500 nucleotides encoding 500 amino acids,in which no inserts and deletions were identified.The identity rates of nucleotide and amino acid in E gene were 99.2％-99.8％ and 100.0％ among the 6 JEV strains isolated from Zhejiang,99.1％-99.3％ and 99.2％-99.8％ between the Zhejiang strains in 2009-2010 and the Zhejiang strains in 2007-2008,respectively,87.6％-88.0％ and 97.8％ between the 6 Zhejiang strains and the vaccine strain SA 14-14-2 of JEV,respectively.The phylogeny tree of E gene indicated that the JEV isolates in Zhejiang during 2009-2010 was located in the branch of the genotype Ⅰ.Conclusion Mosquitoes collected from Cixi and Xianju areas carried JEV,with the rate of JEV in Cixi higher than in Xianju.All the Zhejiang isolates in 2009-2010 were proven to be the genotype Ⅰ of JEV.

OBJECTIVETo describe the experience with extracorporeal membrane oxygenation (ECMO) for cardiorespiratory support of 100 patients.

METHODSRetrospective analysis of the medical files of 100 patients submitted to the implant of extracorporeal membrane oxygenation system for cardiorespiratory assistance of acute and refractory cardiogenic shock from December 2004 to September 2008. There were 67 males and 33 females, age ranged from 5 d to 76 years with a mean of (28+/-26) years, body mass ranged from 3.8 to 100.0 kg with a mean of (42+/-30) kg. The inter-surface of the ECMO equipment system was completely coated by heparin-coating technique. All patients were applied veno-artery ECMO and activated clotting time was maintained between 120 and 180 s and heparin usage dose was 5 to 20 Uxkg(-1)xh(-1). Mean blood flow was 40 to 220 mlxkg(-1)min(-1) during ECMO assistant period.

RESULTSThe shortest ECMO time was 12 to 504 h with a mean of (119+/-80) h. Sixty-one patients (61.0%) weaned off successfully from ECMO, 55 of them (90.2%) were discharged and 6 died of post-operative complications. Thirty-nine patients could not weaned off from ECMO. Total survival discharge rate was 55.0%. Mean aortic pressure before ECMO in survived patients was significantly higher than that of dead patients (P=0.038). Lactic acid concentration of artery blood before ECMO in survived patients was significantly lower than that of dead patients (P=0.005).

CONCLUSIONSECMO is an effective mechanical assistant therapy method for cardiac and pulmonary failure after cardiac surgery. Earlier usage of ECMO for heart lung failure patient and avoiding the main organs from un-recovery trauma are key success.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Extracorporeal Membrane Oxygenation ; Female ; Heart Failure ; therapy ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Respiratory Insufficiency ; therapy ; Retrospective Studies ; Young Adult

OBJECTIVETo investigate the effectiveness of testicular sperm cryopreservation in male fertility preservation by evaluating the clinical outcome of ICSI cycles with frozen-thawed testicular sperm for azoospermia patients.

METHODSWe retrospectively analyzed 96 samples of cryopreserved testicular sperm obtained by testicular biopsy, vasovasostomy (V-V), vasoepididymostomy (V-E) , of which 55 were subjected to 60 ICSI cycles with frozen-thawed testicular sperm. We evaluated the rates of sperm recovery, fertilization, cleavage, transferable and good-quality embryos, clinical pregnancy, pregnancy outcome, and health of the newborns.

RESULTSAll the frozen testicular sperm samples were recovered successfully. The rates of fertilization, 2PN fertilization, cleavage, available embryos and good-quality embryos were 77.6, 69.4, 99.4, 84.5 and 40.8%, respectively. There were transferable embryos in all cycles. Fresh embryos were transferred in 52 of the 60 cycles, with the clinical pregnancy rate of 57.7% (30/52), including 19 singletons and 11 twins, and the rates of implantation and miscarriage were 38.7% (41/106) and 3.33% (1/30). Up to the present time, there have been 20 healthy newborns, including 12 boys and 8 girls, and another 13 ongoing pregnancies. No birth defects have been found so far.

CONCLUSIONDesirable clinical outcomes can be obtained from ICSI cycles with frozen-thawed testicular sperm, and testicular sperm cryopreservation is an effective method of fertility preservation for azoospermia males.

Adult ; Azoospermia ; therapy ; Cryopreservation ; Female ; Fertility Preservation ; methods ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Semen Preservation ; methods ; Sperm Injections, Intracytoplasmic ; methods

Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors (uPAR) in bone marrow fluid and bone marrow tissue from multiple myeloma (MM) patients and assess association of uPAR level with prognosis of MM. Methods uPAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow (control) were examined by ELISA. Furthermore, uPAR expression in bone marrow tissue was investigated by RT-PCR and Western blot, respectively. The distribution of uPAR in MM cells was examined using immunofluorescence staining. The pathological changes in different stages of MM patients were studied by HE staining. Results uPAR level in bone marrow fluid of MM patients (1.52±0.32 μg/ml) was found to be higher than that in the control group (0.98±0.15 μg/ml). Interestingly, uPAR protein (0.686±0.075 vs. 0.372±0.043, P<0.05) and mRNA (2.51±0.46 vs. 4.46±1.15, P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage. The expression of uPAR in MM bone marrow was confirmed by immunofluorescence staining. Moreover, HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma. Conclusion Our study reveals that uPAR expression is positively correlated with the development and progress of MM.

Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors (uPAR) in bone marrow fluid and bone marrow tissue from multiple myeloma (MM) patients and assess association of uPAR level with prognosis of MM. Methods uPAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow (control) were examined by ELISA. Furthermore, uPAR expression in bone marrow tissue was investigated by RT-PCR and Western blot, respectively. The distribution of uPAR in MM cells was examined using immunofluorescence staining. The pathological changes in different stages of MM patients were studied by HE staining. Results uPAR level in bone marrow fluid of MM patients (1.52±0.32 μg/ml) was found to be higher than that in the control group (0.98±0.15 μg/ml). Interestingly, uPAR protein (0.686±0.075 vs. 0.372±0.043, P<0.05) and mRNA (2.51±0.46 vs. 4.46±1.15, P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage. The expression of uPAR in MM bone marrow was confirmed by immunofluorescence staining. Moreover, HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma. Conclusion Our study reveals that uPAR expression is positively correlated with the development and progress of MM.
Adult ; Aged ; Bone Marrow ; chemistry ; Disease Progression ; Female ; Fluorescent Antibody Technique ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; chemistry ; pathology ; Receptors, Urokinase Plasminogen Activator ; analysis