Objective To study the effect of siRNA targeting survivin and tumor necrosis factor related apoptosis-inducing ligand on the proliferation and apoptosis of hepatocellular carcinoma ( HCC) cells. Methods siRNA eukaryotic expression vector was constructed and the effects of soluble tumor necrosis factor related apoptosis-inducing ligand ( sTRAIL) on HCC cells were observed. Results The recombinant plasmid Psilence ( + )-survivin was successfully constructed. Survivin mRNA expression inhibition ratio reached 73% by RT-PCR. Observed by MTT method sTRAIL failed to inhibit the survival rate of HepG2、HepG2/Silence( - ) cells at 12 h、24 h、48 h when compared to control groups. With the survivin gene being inhibited, the survival rate of HepG2/Silence( + ) cells(0. 518?0. 017) decreased in 12 h compared to control groups (0. 741?0. 005 ) and reached the lowest level in 48 h ( P

Aim To study inhibitory effect of 5-fluorouracil encapsulated by galactosylceramide liposomes (5-Fu-GCL)on 5-Fu-resistent HepG_2 cells and its mechanisms. Methods Inhibitory effect of 5-Fu-GCL on established model of 5-Fu-resistant HepG_2 cells was assessed with MTT assay in vitro. The concentration-time course of 5-Fu-GCL in intracellular fluid was detected with high performance liquid chromatography (HPLC). Thymidylic acid synthase (TS) expression was observed with immunohistochemical method,and NO content was determined with chemical method. Results Obvious inhibitory effects of 5-Fu-GCL (75,150,300,600,1200?mol?L-1) on 5-Fu-resistant HepG_2 cells were observed with IC_ 50 of 158.6 ?mol?L-1,far lower than that of free 5-Fu (400.9 ?mol?L-1). 5-Fu-GCL (300 ?mol?L-1) inhibited 5-Fu-resistant HepG_2 cells in a time-dependent manner,and the inhibitory effect of 5-Fu-GCL was stronger than that of free 5-Fu during 12～48 h. Compared with free 5-Fu,5-Fu-GCL (300 ?mol?L-1) increased the content of intracellular fluid in 5-Fu-resistant HepG_2 cells. 5-Fu-GCL(62.5,300,1200 ?mol?L-1) not only inhibited the expression of TS,but also increased the production of NO in 5-Fu-resistant HepG_2 cells,and these effects of 5-Fu-GCL(300,1200 ?mol?L-1) were stronger than those of free 5-Fu. Conclusion 5-Fu-GCL has inhibitory effect on 5-Fu-resistant HepG_2 cells. The effect may be related to the increased concentration of 5-Fu-GCL in intracellular fluid,inhibited expression of TS and increased production of NO.

OBJECTIVETo explore the chaotic dynamic process of multiple organs dysfunction syndrome (MODS) and the regulatory effect of Shenqin Liquid (SQL), a Chinese herbal liquid preparation with the action of purging and qi-tonifying.

METHODSEighty SD rats were divided into 4 groups, and were given suspension of zymosan A and paraffine (1 mL/kg) by peritoneal injection except for those in the blank control group to set up the multiple organs dysfunction syndrome (MODS) model. Low and high doses SQL were administered twice at the doses of 30 and 60 g/kg of SQL respectively at an interval of 8 h per day before modeling. Serum concentration of tumor necrosis factor alpha (TNF-alpha) and nitric oxide (NO) in MODS model animals were tested diachronically, eg. 12, 6 h before modeling, during modeling, 6 and 12 h after modeling, and then the mathematic models were built up with compartment analysis. Lyapunov exponents (LE) of the mathematic models were calculated to evaluate their chaotic characteristics of movement and the degree of chaos was ascertained with the correlation dimension (CD).

RESULTSThe serum levels of TNF-alpha and NO were significantly higher than those in the bland control group at modeling, 6, and 12 h after modeling (P <0.01), while those in the low and high doses of SQL were significantly lower than the model group (P <0.01). Moreover, the level of NO in the high dose of SQL was significantly lower than that in the low dose group (P <0. 01). CD of TNF-alpha movement in the blank control group was 0.803 with the LE less than zero; those in the model group was 1. 966 and > 0 respectively; in the low dose and high dose SQL treated groups, CD was 0.517 and 0.653 respectively and LE >0. CD of NO movement in the blank control group was 0.670 and with LE < 0; in the model group, 1.242 with LE > 0; in the low dose SQL group, 0.574 and in the high dose SQL group 0.850, and LE <0 in the two groups.

CONCLUSIONUnder the normal physiologic condition, TNF-alpha and NO moved steadily without chaotic properties; while under the pathologic condition of MODS, they manifest relatively complicated chaotic properties. SQL can intervene the movement of TNF-alpha and NO, decrease the complexity of their chaotic movement, and make them return back to a stable state.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Multiple Organ Failure ; drug therapy ; metabolism ; Nitric Oxide ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore clinical effects of manipulative reduction with minimally invasive percutaneous plate osteosynthesis in treating distal tibiofibular fractures.

METHODSFrom 2009 to 2011, 60 patients with distal tibiofibular fractures were treated by manipulative reduction with minimally invasive percutaneous plate osteosynthesis. Among them, there were 32 males and 28 females aged from 14 to 70 years old with an average of 41.22 +/- 2.06. According to AO classification of fractures,5 cases were type A1, 22 cases were type A2,21 cases were type A3 and 12 cases were type C1. Operation time, blood loss,time of callus and fracture healing were observed, Mazur scoring of ankle joint were used to evaluate therapeutic.

RESULTSFifty-eight incisions were healed at stage I ,and 2 cases were infected at distal tibial. Operation time was with an average of (62.34 +/- 5.66) min ranged 45 to 90 min;blood loss was 30 to 150 ml with an average of (80.57 +/- 5.59) ml;formation of callus appeared from 4 to 12 weeks,with an average of (8.24 +/- 2.06) weeks, and fracture healing time was from 3 to 6 months, with an average of (4.50 +/- 1.13) months. According to Mazur scoring of ankle joint 40 cases got excellent results, 18 good, and 2 fair.

CONCLUSIONManipulative reduction with minimally invasive percutaneous plate osteosynthesis can obtain reliable fixation. It is a good choice of treating distal tibiofibular fractures by protecting blood supply of fractures.

Adolescent ; Adult ; Aged ; Bone Plates ; Female ; Fracture Fixation, Internal ; instrumentation ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Tibial Fractures ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effect of peroxisome proliferators activated receptors (PPAR) alpha, gamma ligand on ATP-binding cassette transporter A1 (ABCA1) and caveolin-1 expressions and cholesterol, ox-LDL contents in human monocyte derived foam cells.

METHODMalondialdehyde (MDA) was measured by TBARS method, ox-LDL detected by ELISA method, cholesterol measured by fluorescence spectrophotometric method, ABCA1, caveolin-1 mRNA and protein expressions determined by RT-PCR and Western blot, in human monocytes, foam cells [human monocyte-derived macrophage induced by myristate acetate (PMA) further treated with 50 mg/L ox-LDL for 24 h], foam cells plus 10 micromol/L pioglitazone for 48 h, foam cells plus 5 micromol/L clofibrate for 48 h.

RESULTThe intracellular total cholesterol (TC), free cholesterol (FC), cholesteryl ester (CE), ox-LDL and lipid peroxide were significantly increased and the membrane expressions of ABCA1, caveolin-1 were down-regulated in foam cells compared to monocytes (all P < 0.05) and these changes were significantly attenuated by cotreatment with PPARalpha, gamma ligand.

CONCLUSIONThe anti-atherosclerosis effects of PPARalpha, gamma ligand are related to reducing cholesterol contents and up-regulating ABCA1, caveolin-1 expressions in foam cells.

ATP Binding Cassette Transporter 1 ; ATP-Binding Cassette Transporters ; metabolism ; Caveolin 1 ; metabolism ; Cell Line ; Cholesterol ; genetics ; metabolism ; Foam Cells ; metabolism ; Gene Expression ; Humans ; Malondialdehyde ; metabolism ; Monocytes ; metabolism ; PPAR alpha ; metabolism ; PPAR gamma ; metabolism

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

To express recombinant arresten in Escherichia coli (E. Coli) and investigate its biological activity, prokaryotic expression vector of human arresten gene was constructed by gene engineering. Human arresten gene was amplified from recombinant plasmid pGEMArr by polymerase chain reaction (PCR), and inserted into prokaryotic expression vector pRSET containing T7 promoter. Restriction analysis and DNA sequencing verified that the arresten gene was correctly cloned into the expression vector. The recombinant plasmid pRSETAt was subsequently transformed into E. Coli BL21 (DE3), and the target gene was expressed under induction of IPTG. SDS-PAGE analysis revealed that the recombinant protein with a molecular weight of 29 kD (1 kD=0. 992 1 ku) amounted to 29 % of the total bacterial proteins. After purification and renaturation, the recombinant protein could significantly suppress the proliferation of human umbilical vein endothelial cells (HUVECs). These results suggested that the expression of a biologically active form of human arresten in the pRSET expression system laid a foundation for further study on the mechanistic insight into arresten action on angiogenesis and the development of powerful anti-cancer drugs.

Postpartum hemorrhage (PPH) is one of the most adverse obstetric outcomes.Our aim is to detect the risks of multilevel PPH in different cesarean section (CS) groups [including nulliparous CS with indications,nulliparous CS without indications,repeat cesarean (RC),vaginal birth after cesarean (VBAC),cesarean after vaginal birth (CAVB)].We conducted a retrospective cohort study,and the data on 127 145 women collected from January 2014 to May 2016 and from 35 tertiary hospitals in Shanxi province,China,were reviewed.Based on the measuring results of PPH,an ordered logistic regression model was used to analyze the adjusted PPH risks for each of the CS groups,and comparisons were drawn between them.Finally,a total of 99 066 nulliparous (77.92％) and 28 079 multiparous (22.08％) women were observed.The number of CS cases was 61 117,and the rate for CS was 48.07％.A total of 10 029 women did not show indications for CS and accounted for 16.41％ of the CS parturient,whereas 9103 women underwent a repeated cesarean,with a CS frequency of 14.89％.The number of VBAC cases was 989,whose rate was 9.88％ in prior CS women.The number (proportions) of PPH was 3658 (2.88％) in LI (PPH volume:≥900 and ＜1500 mL),520 (0.41％) in L2 (PPH volume:≥1500 and＜2100 mL),and 201 (0.16％) in L3 (PPH volume:≥2100 mL).The Ln (n=1,2,3,etc.) represented the increasing order of PPH severity.In the adjusted results,compared with spontaneous vaginal delivery (SVD) as the reference group,in the adjusted result for nulliparous,there was a decreased PPH risk in CS with indications (OR:2.32;CI:2.04-2.62),which was lower than that of CS without indications (OR:2.50;CI:2.01-2.96).The highest PPH risk in all subgroups (i.e.nulliparous and multiparous groups) was observed in the RC (OR:3.61;CI:3.16-4.17),which was nearly twice higher than that of the VBAC (OR:1.82;CI:1.33-2.52).CAVB (OR:1.03;CI:0.65-1.62) showed no significant difference with the reference group.Thus,we deemed that CS should be avoided in nulliparous pregnancies unless indicated,to prevent or reduce the rates for the use of RC or VBAC which are high risks of severe PPH to the parturient women.

Background: Advances in blood biomarker discovery have enabled the improved diagnosis and prognosis of Alzheimer's disease (AD). Most branched-chain amino acids, except isoleucine (Ile), are correlated with both mild cognitive impairment (MCI) and AD. Therefore, this study investigated the association between serum Ile levels and MCI/AD. Methods: This study stratified 700 participants from the Alzheimer's Disease Neuroimaging Initiative database into four diagnostic groups: cognitively normal, stable MCI, progressive MCI, and AD. Analysis of covariance and chi-square analyses were used to test the demographic data. Receiver operating curve analyses were used to calculate the diagnostic accuracy of different biomarkers and were compared by MedCalc 20. Additionally, Cox proportional hazards models were used to measure the ability of serum Ile levels to predict disease conversion. Finally, a linear mixed-effects model was used to evaluate the associations between serum Ile levels and cognition, brain structure, and metabolism. Results: Serum Ile concentration was decreased in AD and demonstrated significant diagnostic efficacy. The combination of serum Ile and cerebrospinal fluid (CSF) phosphorylated tau (P-tau) improved the diagnostic accuracy in AD compared to total tau (T-tau) alone. Serum Ile levels significantly predicted the conversion from MCI to AD (cutoff value of 78.3 μM). Finally, the results of this study also revealed a correlation between serum Ile levels and the Alzheimer's Disease Assessment Scale cognitive subscale Q4. Conclusions Serum Ile may be a potential biomarker of AD. Ile had independent diagnostic efficacy and significantly improved the diagnostic accuracy of CSF P-tau in AD. MCI patients with a lower serum Ile level had a higher risk of progression to AD and a worse cognition assessment.

Background: Advances in blood biomarker discovery have enabled the improved diagnosis and prognosis of Alzheimer's disease (AD). Most branched-chain amino acids, except isoleucine (Ile), are correlated with both mild cognitive impairment (MCI) and AD. Therefore, this study investigated the association between serum Ile levels and MCI/AD. Methods: This study stratified 700 participants from the Alzheimer's Disease Neuroimaging Initiative database into four diagnostic groups: cognitively normal, stable MCI, progressive MCI, and AD. Analysis of covariance and chi-square analyses were used to test the demographic data. Receiver operating curve analyses were used to calculate the diagnostic accuracy of different biomarkers and were compared by MedCalc 20. Additionally, Cox proportional hazards models were used to measure the ability of serum Ile levels to predict disease conversion. Finally, a linear mixed-effects model was used to evaluate the associations between serum Ile levels and cognition, brain structure, and metabolism. Results: Serum Ile concentration was decreased in AD and demonstrated significant diagnostic efficacy. The combination of serum Ile and cerebrospinal fluid (CSF) phosphorylated tau (P-tau) improved the diagnostic accuracy in AD compared to total tau (T-tau) alone. Serum Ile levels significantly predicted the conversion from MCI to AD (cutoff value of 78.3 μM). Finally, the results of this study also revealed a correlation between serum Ile levels and the Alzheimer's Disease Assessment Scale cognitive subscale Q4. Conclusions Serum Ile may be a potential biomarker of AD. Ile had independent diagnostic efficacy and significantly improved the diagnostic accuracy of CSF P-tau in AD. MCI patients with a lower serum Ile level had a higher risk of progression to AD and a worse cognition assessment.