Objective To investigate the effects of ubiquitin isopeptidase inhibitor Ⅰ on proliferation and apoptosis of human leukemic K562 cells.Methods Human leukemic K562 cells were treated with different concentration of ubiquitin isopeptidase inhibitor Ⅰ.Cell proliferation was monitored by MTT assay.Cell apoptosis was analyzed by flow cytometry after Annexin V/PI staining,and mRNA expression levels of bcl-2,bax and Caspase-3 were quantified by reverse transcription-polymerase chain reaction (RT-PCR).Results MTT assay showed that the proliferation of K562 cells was markedly inhibited by ubiquitin isopeptidase inhibitor Ⅰ in a time-and-dose dependent manner.The Annexin V positive rate of K562 cells after treatment with ubiquitin isopeptidase inhibitor Ⅰ was significantly increased by FCM analysis (P ＜ 0.05).The early apoptosis rate of K562 cells treated with 100 nmol/L ubiquitin isopeptidas inhibitor Ⅰ by 72 h,was obviously increased compared to control cells [(15.4±1.3) ％ vs (4.1±0.9) ％,P ＜ 0.01].The mRNA levels of Caspase-3 and bax were up-regulated and bcl-2 was down-regulated after treated with ubiquitin isopeptidase inhibitor Ⅰ,and the differences were statistically significant from control cells (P ＜ 0.05).Conclusion Ubiquitin isopeptidase inhibitor Ⅰ has inhibitory effect on proliferation and inductive effect on apoptosis of K562 leukemia cells,implying its possible application in treating leukemia.

0 05) The maternal plasma CGRP level was significantly higher(P

Objective To investigate the mechanism involved in the brain protection afforded by prenatal hypoxic adaptation by determining the quantitative variation in bcl-2 and bax mRNA expression.Methods Twenty-four Wistar pregnant (22d pregnant) rat were randomly divided into two groups: group I (hypoxia group) and group *** ( control group) . In group Ⅰ the pregnant rats were placed in an airtight cabin specially designed for hypoxic adaptation. When O2 % in the cabin decreased to 15%, the animals were taken out breathing fresh air for 5min and then placed back in the cabin and underwent another episode of hypoxia. In group Ⅱ the animals were placed in the cabin which was not tightly closed and underwent no hypoxia. 7 fetal or newborn rats were taken at 1st, 3rd, 24th, 48th, 72nd, 120th, and 168th h after prenatal hypoxic adaptation from each group and their brains removed for determination of bcl-2 mRNA and bax mRNA. Results In control group the expression of bcl-2 and bax were observed in the brain tissue of normal fetal or newborn rats from the 22nd day in the uterus to the 7th day postpartum during which there were no significant changes in bcl-2 gene expression while bax gene expression gradually decreased with time ( the decrease was of no statistical significance) . In hypoxia group bax gene expression decreased at 8th h after hypoxic adaptation and reached the bottom at 24th h which persisted until 120th h; while bcl-2 gene expression started increasing at 24th h after hypoxic adaptation and persisted until 72nd h. The bcl-2/bax ratio also started increasing at 8th h after hypoxic adaptation and peaked at 24th h and persisted until 72nd h. Conclusions In the brain tissue of fetal and newborn rats which have undergone prenatal hypoxic adaptation, bcl-2 gene expression is elevated, bax gene expression decreases and bcl-2/bax ratio increases. These changes are time -dependent.

Acetyl-CoA C-acetyltransferase (AACT) is the first enzyme in the terpene synthesis pathway, catalyzed two units of acetyl-CoA to acetoacetyl-CoA. In order to study the tanshinone biosynthesis in Salvia miltiorrhiza, a novel AACT gene, SmAACT, was cloned using cDNA microarray and RACE strategy. The full length cDNA of SmAACT is 1 623 bp (accession No. EF635969), which contained a 1 200 bp open reading frame (ORF) encoding a 399 amino acid protein. Nine introns were found in the genomic sequence. SmAACT was upregulated by YE and Ag+ elicitors both with cDNA microarray and quantitative RT-PCR analyses along with the accumulation of tanshinones. Sequence homology comparison and phylogenetic analysis all suggested that SmAACT belonged to the class of acetyl-CoA C-acetyltransferase. The transcription level of SmAACT was relatively higher in root than that in stem and leaf tissues. SNP analysis revealed that SmAACT was highly variable in the region of 6 to 9 introns with 33 SNPs in the 600 bp region, there are 5 SNPs in the cDNA region while they are all synonymous cSNPs. Some special genotypes were found in Salvia miltiorrhiza from different areas. SmAACT will be an useful gene for further analyze the mechanism of gene regulation among the tanshinones biosynthesis.

Objective To investigate the clinical features and causes of micturition and defecation dysfunction in motor neuron disease (MND) patients.Methods The micturition and defecation function was evaluated by a questionnaire covering storage and voiding of urine and feces respectively in 50 MND patients.The clinical features and external anal sphincter electromyography (EAS-EMG) were analyzed to explore the causes of micturition and defecation dysfunction in MND patients.Results Micturition and defecation dysfunction was detected in 9 of 50 (18.0％) MND patients.The main types of micturition and defecation dysfunction were constipation (4/9),urinary frequency,urgency with or without incontinence,fecal urgency (4/9),powerlessness for micturition and defecation (2/9),hesitancy for micturition (1/9).EAS-EMG was normal in 9 MND patients accompanied with micturition and defecation dysfunction.Conclusions MND patients accompanied with micturition and defecation dysfunction were not very rare.Constipation,urgency and powerlessness were the main types of micturition and defecation dysfunction and they were not related to the function of external anal sphincter.Gastrointestinal dysfunction from abnormal autonomic nerve involvement,muscle weakness and the resulted reduced activity,severe upper motor neuron damage and respiratory muscle weakness may be the main causes of micturition and defecation dysfunction in MND patients.

Objective To explore the impact of childbirth on women external anal sphincter electromyography by analysis of motor unit potentials parameters of external anal sphincter electromyography of healthy women volunteers with different childbirth experiences.Methods Twenty-eight healthy volunteers with different childbirth experiences were recruited in this study.Among them , 7 had a history of cesarean and 21 had experienced 1-3 uncomplicated vaginal deliveries ( 12 women had experienced 1, 6 had experienced 2, and 3 had experienced 3).Another 9 nulliparous healthy volunteers were recruited as control group.Among all subjects who completed the examination , parameters of motor unit potentials ( duration, amplitude, area, mean number of turns and mean phases ) of bilateral external anal sphincter were obtained for statistical analysis.Results There were no significant differences in any of the parameters of motor unit potentials between the nulliparous group and the cesarean group , while parameters of durations (10.5 ms vs 9.0 ms, t=-2.506, P=0.019) and mean phases (4.0 vs 3.6, t =-2.707, P=0.012) of vaginal delivery group were significantly higher than that of cesarean group and parameter of durations ( 10.5 ms vs 8.9 ms, t =-3.025, P =0.005 ) of vaginal delivery group was significantly higher than that of the nulliparous group;With increasing numbers of vaginal deliveries , only the parameter of mean turns showed statistically significant differences , although there was a slight trend of larger and more complex motor unit potentials.Compared with subjects without vaginal delivery experience ( including nulliparous group and the cesarean group ) , the parameters of durations and mean phases of subjects with vaginal delivery experience were statistically higher; Nevertheless , parameter of bilateral duration of the motor unit potential preferred asymmetric.Conclusions Childbirth experience has an explicit impact on the parameters of the external anal sphincter motor unit potentials , which is considered to be strongly correlated with the vaginal delivery process rather than the pregnancy procession itself.In addition , the impact is mostly from the first vaginal delivery experience.The characters of asymmetry of motor unit potential parameters imply the possibility of mechanical damage and decrease blood perfusion of tissues during delivery procession.

Objective The feasibility and the curative effect of bipolar hemiarthroplasty for hemiplegic side femoral neck fractures in elderly patients were evaluated.Methods The patients suffering from hemiplegic side femoral neck fractures and undergoing bipolar hemiarthroplasty in Beijing Tongren Hospital were followed-up during 2012-2015.The postoperative Harris Score and the complications such as infection,deep vein thrombosis and dislocation were collected.Results Totally 25 patients(25 hips)were followed-up.The average age was 70.1 years,and mean follow-up period was 24.3 months (10-42 months).The mean Barthel index score was higher (77.4 ± 14.7,ranging from 40-95 points) after operation than before operation[25.00 ± 5.95 (15-35 points)] with a statistically significant difference(t=-21.374,P＜＜0.05).The postoperative Harris Score was 83.5± 8.9(60-95 points).According to Harris score,the excellent and good rate was 80.0％.The patient's satisfaction rate was 88.0％.Conclusions Choosing proper prosthesis and fixing measure,and keeping the function and tension of short extorsion muscles can achieve the satisfactory results for hemiplegic side femoral neck fractures in elderly patients.

The RAS (rat sarcoma) gene is one of the important oncogenes, and its mutation is present in about 30% human tumors. KRAS (kirsten rat sarcoma viral oncogene) is one of the three RAS subtypes, and KRAS mutations are more common than the mutations in other two RAS subtypes. In recent years, with the continuous research, new ideas have been provided for the treatment of cancers via targeting-KRAS. Efforts have been made to develop various KRAS inhibitors. Here, based on the mechanism of action, we classified KRAS inhibitors into two categories: inhibitors that directly target KRAS and inhibitors that indirectly act on KRAS. The representative KRAS inhibitors were summarized and introduced in this paper.

Objective To detect the expression and activity of AMP-activated protein kinase α subunit (AMPKα) and peroxisome proliferator-activated receptor-α (PPARα) in liver of high-fat fed rats treated with metformin, and to investigate the mechanisms underlying metformin decreasing the total cholesterol (TC) and triglyceride (TG) contents of the liver. Methods Total 30 male Wistar rats were randomly divided into control group (group C), high-fat diet fed group (group HF) and high-fat diet feeding plus metformin treatment group (group Met,metformin was administered orally at the last month of high-fat diet feeding). After feeding for 5 months, TC and TG in liver and sera were determined, respectively. The mRNA and protein levels and activity of AMPKα and PPARα in the liver were determined by real-time PCR and Western blotting. The activity of PPARα transcriptor binding to DNA was detected by ELISA. Results Five months of high-fat diet feeding induced a significant decrease in AMPKα and phosphorylated-AMPKα protein expression as well as AMPKα2 and PPARα mRNA levels in the liver of rats (all P＜0.05), while it did not alter PPARα protein expresssion and the PPARα activity binding to DNA as well as AMPKα1 mRNA levels. The TC and TG contents in the liver (P＜0.05) and serum (P＜0.05) were sharply increased in group HF than those in group C. Treatment with metformin for 1 month led to a marked increase of AMPKα2 mRNA level, AMPKα and phosphorylated-AMPKα protein expression as well as the PPARα activity in group Met compared with group HF(all P＜0.05), while the PPARα protein expression and the PPARα mRNA level did not show significant change. Consistent with these findings, the TC and TG contents in rat liver as well as sera were strikingly decreased (all P＜0.05). Conclusion The activations of AMPKα and PPARα induced by metformin may contribute to the decrease of TC and TG content in liver and sera of the high-fat fed rats.

Objective To explore effect of physic temperature dropping by ice blanket on incidence of pressure ulcer of stroke patients in ICU, and normalize the basic nursing procedure during temperature reduc-tion.Methods 230 fever patients were randomly divided into the group A(control temperature with ice blan-ket,108 cases)and the group B (control temperature with routine measure,122 cases).Thc incidence of pressure ulcer and surface temperature of pressed position in the two groups were compared.Results The incidence of pressure ulcer and surface temperature of pressed position in group A was lower than those of group B.Conclusions The incidence of pressure ulcer can be reduced by using ice blanket for physic temperature dropping.The pressure of pressed position were descended by alternative prostration and lateral position.