Objective To explore effects of two intravenous infusion methods on elderly patients.Methods Self-control method was used on 568 cases of elderly patients.Traditional method was used on Day 1:The adjustable clamp was fully opened when a free flow of blood appeared during venipuncture.The experimental method was used on Day 2:The adjustable clamp was opened 1/3 when a free flow of blood appeared during venipuncture.Success rate of venipuncture and value of pain were compared between the two groups.Results The experimental method had a higher success rate of venipuncture than the tranditional method (97.89％ vs 93.31％,x2 =13.098,P ＜ 0.01 ).There was no significant difference in the mean value of pain during a successful venipuncture ( P ＞ 0.05 ).During an unsuccessful venipuncture,mean value of pain in the experimental group was lower than that in the traditional method group,and the difference was statistically significant [ ( 1.97 ±0.52) vs (2.35 ± 0.66 ),t =4.987,P ＜ 0.01 ].Conclusions Opening the adjustable clamp for 1/3 when a free flow of blood appeared during venipuncture on elderly patients could improve the success rate of venipuncture and decrease pain during an unsuccessful venipuncture.

AIM:To investigate the effect of small interfering RNA(siRNA)-mediated progranulin(PGRN) gene silencing on the proliferation and migration abilities of human non-small-cell lung cancer A549 cells and its mecha-nism.METHODS:The mRNA and protein expression levels of PGRN in the A 549 cells and human bronchial epithelial (HBE)cells were detected by qPCR and Western blot.A549 cells were transfected with PGRN-siRNA by liposome meth-od.The expression of PGRN at mRNA and protein levels in the A 549 cells transfected with PGRN-siRNA was detected by qPCR and Western blot, respectively.The cell viability was measured by MTT assay.The cell proliferation ability was measured by living cells counting and crystal violet staining assays.The cell migration ability was measured by wound-heal-ing and Transwell assays.The protein levels of proliferating cell nuclear antigen(PCNA),cyclin D1,Bcl-2 and Bax were determined by Western blot.The protein levels of phosphorylated extracellular signal-regulated kinase 1/2(p-ERK1/2) and phosphorylated protein kinase B(p-Akt)were also determined by Western blot.RESULTS:The expression of PGRN at mRNA and protein levels was higher in the A 549 cells than that in the HBE cells(P<0.05).The expression of PGRN at mRNA and protein levels in the A549 cells transfected with PGRN-siRNA was significantly decreased,and the cell pro-liferation and migration abilities were significantly decreased.The protein expression levels of PCNA,cyclin D1 and Bcl-2 were significantly reduced and the protein expression level of Bax was significantly increased(P<0.05).Meanwhile,the protein levels of p-ERK1/2 and p-Akt were down-regulated(P<0.05).CONCLUSION:PGRN gene silencing obviously inhibits the proliferation and migration abilities of human non-small-cell lung cancer A549 cells.The PI3K/Akt and MAPK/ERK signaling pathways may play an important role in these processes.

Coronary artery disease (CAD) is one of the leading causes of death. Safflower attracts great attention owing to its anti-ischemia/reperfusion injury effect. Ninety-three patients with CAD were included and randomized into safflower treatment group, PCI group and control group. Low-dose dobutamine stress echocardiography (DSE) was performed to measure end-systolic volume (ESV), end-diastolic volume (EDV), left ventricular ejection fraction (LVEF) and wall motion score index (WMSI) to determine the recovery of hibernating myocardium and cardiac function in all patients before treatment and after 3-month follow-up. The study was to investigate the effects of safflower on hibernating myocardial revascularization and cardiac function. It was found that LVEF was significantly improved, while the ESV and WMSI were significantly reduced after 2-week treatment in safflower and PCI treatment groups. No significant differences were found between safflower and PCI treatment groups in ESV, EDV, WMSI and LVEF after treatment Safflower injection effectively improved hibernating myocardial function.
Aged ; Carthamus tinctorius ; chemistry ; Coronary Artery Disease ; drug therapy ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Heart ; drug effects ; physiopathology ; Humans ; Male ; Middle Aged ; Myocardial Revascularization ; Myocardial Stunning ; drug therapy ; physiopathology ; surgery ; Recovery of Function

Apoptosis of abnormal oocytes is essential for defective oocyte elimination during prepubertal ovary development, and the ubiquitin system regulates the cell apoptosis via the degradation of specific proteins. Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a component of the ubiquitin system, and the UCH-L1-dependent apoptosis is important for spermatogenesis. In the present study, the change in the number of follicles and the expression of UCH-L1 in oocytes were determined in prepubertal mouse ovaries by immunohistochemical techniques. A significant decrease in the follicular pool was found in prepubertal mouse ovaries during the period of day 21 to day 28 after birth, and accordingly, the UCH-L1 protein expression was increased, to some degree in association with Jun activation domain-binding protein 1 (Jab1) and cyclin-dependent kinase inhibitor p27(Kipl). The increased UCH-L1 protein, together with the corresponding changes of Jab1 was detected in morphologically abnormal oocytes of prepubertal ovaries. Through the immunofluorescent colocalization, UCH-L1 was shown concentrating in abnormal oocytes, and a parallel change in Jab1 was also seen. The affinity analysis confirmed the interaction between UCH-L1 and Jab1 in ovaries. These results suggest that UCH-L1 plays an important role, possibly in association with Jab1 and p27(Kipl), in selective elimination of abnormal oocytes during mouse prepubertal development.
Animals ; Apoptosis ; Female ; Mice ; Oocytes ; cytology ; Ovary ; enzymology ; Ubiquitin Thiolesterase ; metabolism

OBJECTIVETo construct four expression vectors carrying enhanced green fluorescent protein (EGFP) gene under the control of different HBV promoters, and to detect and analyze their expressions in hepatoma cell lines.

METHODSFour HBV promoters were amplified using PCR, and they were inserted into the T-vector and identified using restriction enzymes and sequencing, then cloned into the expression vector pEGFP-1. The four recombinant plasmids were transfected into human hepatoma cell line HepG2 by lipofectamine2000, and the positive cell clones were detected using fluorescence microscopy.

RESULTSAll target fragments were separately obtained and successfully cloned into the expression vector. The expressions of EGFP under the control of the four promoters were detected. The expressions of EGFP controlled by different promoters had some differences.

CONCLUSIONSReporter gene EGFP under the control of four HBV promoters can be specifically expressed in hepatoma cell line HepG2, and different promoters give different results; this may provide another option in gene therapy of liver diseases.

Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Eukaryotic Cells ; metabolism ; Genetic Therapy ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HeLa Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; pathology ; Promoter Regions, Genetic ; genetics ; Transfection

OBJECTIVE: To explore the effect of precursor of brain-derived neurotrophic factor (proBDNF) on cultured hippocampal neuron and its intracellular mechanism. METHODS: The hippocampal neurons were dissociated from E18 rats and cultured in neurobasal medium, and then the cells were treated with proBDNF, preBDNF(propeptide of proBDNF) and proBDNF antiserum,respectively. Thirty minutes, 1 hour or 48 hours later, the cells were stained with Nissl solution, and the immunocytochemistry methods of ELK-p (Ets domain protein), ErK2(extracellular signal regulated kinase) and c-fos were performed. RESULTS: The expressions of ELK-p, ErK and c-fos were significantly upregulated in the cultured hippocampal neurons after they were treated with proBDNF protein,and the immuno-positive staining was also obvious in some nuclei. While the endogenous proBDNF was neutralized by proBDNF antiserum treatment, the expressions of ELK-p, ErK and c-fos were downregulated and many cells showed swelling and vasoculation. The immunoreactivity in preBDNF treated cells was similar to that in normal cultured cells. CONCLUSION proBDNF plays an important role in sustaining the hippocampal neuron survival through upregulating the ELK and ErK pathways.
Animals ; Brain-Derived Neurotrophic Factor ; pharmacology ; Cell Survival ; drug effects ; Cells, Cultured ; Hippocampus ; cytology ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Neurons ; cytology ; metabolism ; Protein Precursors ; pharmacology ; Proto-Oncogene Proteins c-ets ; metabolism ; Proto-Oncogene Proteins c-fos ; metabolism ; Rats ; Signal Transduction

Objective To investigate the efficacy of evidence-based nursing for prevention of pressure sores in senile orthopedics patients. Methods There were 326 senile orthopedics patients received evidencebased nursing in the observation group, and 268 senile orthopedics patients received usual care belonged to control group. To calculate and compare the incidence rate of pressure sores of the two groups. Results The incidence rate of pressure sores in observation group was 0.7% but 4. 2% in control group, the difference between the two groups had statistical significance (P<0. 05). Conclusions Evidence-based nursing can change clinical nurses'habits and behaviors which based on experience. Nursing practice will be promoted, and senile patient's pressure sores will be prevented effectively through using scientific evidence-based nursing theory to guide clinic

Objective To estimate correlation between phonetically balanced maximum(PB max)and pure tone auditory threshold in auditory neuropathy(AN)patients.nethods 0ne hundred and six ANpatients were identified using multipie criteria including PB max,a metric for speech recognition,pure tone auditory threshold.acoustic emission test.distortion products otoacoustic emission(DPOAE) and auditory brainstem response(ABR).SPSS statistical software was used to estimate the Pearson's correlation between PB max and pure tone auditory threshold and to test whether pure tone auditory threshold,or auditory configuration had a significant impact on PB max.Results Even the patients had the same or similar values for pure tone auditory threshold or auditory configuration.varied values of PB max were found in two hundreds and twelve ears for 106 patients.Analysis of the data for 106 patients revealed a negative correlation(r=-0.602,P<0.01) between PB max and pure tone auditory threshold,i.e.hearing loss at a mild relates to a lower PB max.By using analysis of variance(ANOVA)method,it Was found that both pure tone auditory threshold and auditory configuration had a significant(P<0.01)impact on the patients' PB max.Conclusions This analysis implicated the promise and potential of pure tone auditory threshold and auditory configuration for predicting PB max of the AN patients,and improving the diagnosis of AN.

To investigate the effect of hepatitis B virus X protein(HBx) on CtBP-interacting protein(CtIP) which is an important repair factor of DNA double strand break damage in HepG2 cells induced by bleomycin. A HBx stably expressing HepG2 cell line and a control HepG2 cell line with empty vector transfected were established. After the double strand break (DSB) damage occurred, the mRNA and protein levels of CtIP were detected by Real-time PCR and Western blot assay respectively, cell cycle profiles and apoptotic cell death were determined by a flow cytometry, and the position of CtIP in cells was observed by confocal laser scanning microscopy. It showed that HepG2 cells transfected with hepatitis B virus X gene could stably express HBx protein. After being induced by bleomycin, the percentage of apoptotic cell was 16.90%+/-0.89% in HBx stably expressing HepG2 cell line and 15.30%+/-0.86% in control cell line, respectively (q = 2.074, P is more than to 0.05). While the percentage of death cell was 8.71%+/-0.74% in HBx stably expressing HepG2 cell line and 4.90%+/-0.46% in control cell line, respectively (q = 7.126, P is less than to 0.01). The two cell lines manifested the increase of G2/M arrest and significant difference existed between the two cell lines. HBx down regulated the expression levels of CtIP and its mRNA. The CtIP level was 0.66+/-0.04 in HepG2-HBx cell and 0.73+/-0.05 in HepG2-vec cell, respectively (t = 2.314, P is less than to 0.05). The relative mRNA level was 1.00+/-0.06 in HepG2-HBx cell and 1.23+/-0.08 in HepG2-vec cell, respectively (t = 2. 732, P is less than to 0.05). We also found that CtIP was concentrated in the cell nucleus. The research suggests that HBx may affect DNA-repair pathways by disrupting the expression of CtIP.
Carcinoma, Hepatocellular ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; metabolism ; Real-Time Polymerase Chain Reaction

This study was aimed to clarify whether valproic acid (VPA) induces apoptosis of leukemia HL-60 cell line and its possible mechanism. The effect of different concentrations and treatment time of VPA on HL-60 cell proliferation was assayed by cytotoxicity test (CCK-8 method) and fluorescence microscopy, and flow cytometry was used to detect cell apoptosis. The expressions of telomerase subunit h-tert mRNA and apoptosis-related protein as well as caspase-3 activity were detected by real time-quantitative PCR, Western blot and ELISA respectively. The results indicated that VPA inhibited proliferation of HL-60 cells and induced cell apoptosis in a dose dependent manner (r = -0.87). The expressions of anti-apoptotic protein BCL-2 and h-tert mRNA were significantly decreased while the pro-apoptotic protein BAX and caspase-3 activity increased after treatment with VPA. The apoptosis rate of HL-60 cell was negatively correlated with expression of h-tert mRNA. It is concluded that VPA can inhibit leukemia HL-60 cell proliferation and induce apoptosis. The VPA displays anti-leukemia activity possibly through reducing h-tert mRNA and BCL-2 protein expression, increasing BAX expression and activity of caspase-3.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; HL-60 Cells ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Telomerase ; metabolism ; Valproic Acid ; pharmacology ; bcl-2-Associated X Protein ; metabolism