Objective To construct a recombinant interfering RNA(miRNA)plasmid vector targeting annexinA3 and observe its impact on the growth and apoptosis of human gallbladder cancer cell line SGC-996.Methods Four small fragments of miRNA and one negative control were sequenced and cloned into vector pcDNATM 6.2-GW/EmGFPmiR.The recombinant plasmids were then transfected into gallbladder cancer cell line SGC-996 by positive liposomal transfection.The transcription and translation level of annexinA3 expression were detected by RT-PCR and Western Blot.Meanwhile, the proliferation and apoptosis of transfected tumor cells were determined by MTT and flow cytometry.Results The recombinant plasmids containing annexinA3 miRNA were successfully constructed.AnnexinA3 expression was significantly knocked down after miRNA plasmid transfection.Transfection of annexinA3 gene miRNA could effectively inhibit the proliferation of gallbladder cancer cell and promote apoptosis.Conclusion AnnexinA3 gene silenced by miRNA can induce the apoptosis of human gallbladder cancer cell line SGC-996.The process might serve as a potential approach for cancer gene therapy.

Background and purpose:Radiofrequency ablation (RFA) was one of the best combined treatments for hepatocarcinoma. CT was commonly used to evaluate treatment response and recurrent disease. However it was diffi cult to evaluate treatment effectiveness of RFA and to detect recurrent nodule especially the size below 1cm. Digital subtraction arteriography(DSA) may provide fi nal diagnosis of recurrent nodule after RFA for hepatocarcinoma. Our purpose was to analyze the signs of DSA after RFA for hepatocarcinoma in order to provide possible reference for treatment effectiveness and the imaging follow-up methods. Methods:17 patients with primary liver cancer (n=15) or hepatic metastasis(n=2) were enrolled in this study. Common hepatic arteriography or super-selective angiography of suspicious tumor area were performed on all patients. Results:On DSA, most ablated regions presented as round or ovoid low density areas with no stain. At the peripheral zone of the lesion, fi ve signs after RFA could be found: Stain of localized granulation tissue, arterio-portal fi stula, hemorrhage, recurrence and no abnormal fi ndings. Local or intrahepatic recurrence occurred in 9 cases. Conclusion:Hepatic DSA has great value in detecting local recurrence of hepatic tumor after RFA treatment. DSA is superior to CT in detecting marginal or intrahepatic recurrent nodule below 1 cm.

0.4, P

Objective To compare the effects of no-rinse skin cleanser and warm water with paraffin oil in first brush bath of the new-born. Methods About 42 newborns who were born from January to March 2016 were set as control group and used warm water with paraffin oil in first brush bath. About 43 newborns who were born from April to June 2016 were set as observation group and used no-rinse skin cleanser in first brush bath. The cleaning time and effect ,the rectal temperature before and after sponge and dermatitis were compared. Results Both groups did not develop dermatitis. But the observation group had better cleaning satisfaction , less cleaning time, faster rewarming than those of the control group with statistical significance (all p<0.05). Conclusion No-rinse skin cleanser used in the first brush for the new-born can reduce cleaning time and difference of temperature, which is safe for clinical application.

Objective:To explore the expression of R-spondin family in colorectal cancer tissues and adjacent tissues,and to evaluate its relationship with clinic-pathological stage.Methods:A total of 64 samples of colorectal cancer tissues and adjacent tissues were collected from the patients,who received radical surgery in Xiangya Hospital,Central South University between January 2014 and August 2014.The mRNA and protein expression levels of R-spondin 1-4 and β-catenin in the colorectal cancer tissues and adjacent tissues were detected by qRT-PCR and immunohistochemistry.The relationship between the expression level of R-spondin 1-4 and the clinic-pathological factors were analyzed to explore the correlation between the expression level of R-spondin 1-4 and β-catenin in colorectal cancer.Results:Compared with the adjacent tissues,the mRNA and protein expression levels of R-spondin 1 were elevated in the colorectal cancer tissues (P＜0.05).The mRNA and protein expression levels of R-spondin 2-4 were increased in the colorectal cancer tissues than those in the normal tissues (P＜0.05),but there was no significant difference between the colorectal cancer tissues and adjacent tissues (P＞0.05).The expression level of R-spondin i was positively correlated with the nuclear expression of β-catenin in the colorectal cancer tissues (r=0.6307,P＜0.05).Conclusion:Compared with the adjacent tissues,the mRNA and protein expression levels of R-spondin 1 are significantly elevated in the colorectal cancer tissues.R-spondin 1 may play a role in promoting carcinogenesis by regulating the activity of β-catenin in the downstream of Wnt signaling pathway.

Objective To investigate the causes, clinical features, classifications and liver function change of drug-induced liver damage (DILD) in the elderly. Methods One hundred and sixty seven inpatients with acute drug-induced liver injury in our hospital in the past ten years (January 2000 to December 2009) were retrospectively investigated,and the diagnosis and classification methods of acute DILD were based on international consensus meeting (international criteria). Results Among 167 DILI cases, there were 53 cases (31.7%) in the older group and 114 cases (68.3%) in middle-youth age group. Fatigue and jaundice were the more common symptoms, accounting for 50.3% and 46.7%, respectively. In 167 cases, no obvious symptoms and signs were shown in 25 cases. There were no significance differences in clinical manifestation between two groups. Many drugs could induce liver injury. The most common drugs inducing DILD were Chinese traditional and herbal drugs, accounting for 47.9%. Drugs used in heart diseases and inducing liver injury were more common in the older group. In this study, 40 (75.5%), 5 (9.4%) and 8 (15.1%) cases were designated as hepatocellular, cholestetic and mixed DILD in the older group, and 91 (78.9%), 8 (7.4%) and 15 (13.7%) in middle-youth age group, respectively. There were no significance differences between two groups in classifications. Conclusions Many drugs could cause liver injury. The symptoms of acute DILD are nonspecific. Drugs used in heart diseases and inducing liver injury are more common in older patients.

Objective To discuss the surgical indications,relative merits and clinical outcomes of anterior reconstruction plate fixation for treating unstable posterior pelvic ring injuries.MethodsA retrospective study was done on clinical data of 36 patients with unstable posterior pelvic ring injuries treated with anterior reconstruction plate fixation from August 2004 to July 2010 and followed up for minimum one year.There were 25 males and 11 females,at mean age of 35.8 years ( range,13-62 years).There were two patients with anterior dislocation of the sacroiliac joint and 34 with posterior dislocation.According to Tile classification,there were four patients with type B fractures ( B1:3,B2:1 ) and 32 with type C fractures (C1-1:17,C1-2:12,C2:2,C3:1 ).Results All patients were followed up for average 2.3 years ( range,1-6 years).Fat liquefaction and superficial infection were founded in two patients who were cured by dressing change and frequent but low-dose blood transfusion.Eight patients were subjected to iatrogenic lateral femoral cutaneous nerve injury.Meanwhile,five of them was recovered after treatment with neurotrophic drugs,but there were still three patients leaving lateral thigh numbness.One patient had L5 nerve root injury due to pull force,and was recovered after three months of neurotrophic drug therapy.According to Matta and Tometta criteria,the reduction results were excellent in 23 patients,good in 11 and fair in four,with excellence rate of 94％.No reduction loss or implant failure occurred.According to Majeed scoring system,the clinical outcomes were excellent in 17 patients,good in 14,fair in four and poor in one,with excellence rate of 86％.Of the seven patients pre-operatively associated with sacral plexusinjury,three obtained full recovery,two got partial recovery and two were free from recovery.Conclusion The anterior reconstruction plate fixation takes advantages of wide surgical indications,simple exposure,low infection rate,satisfactory reduction and solid fixation and may be a main treatment method for unstable posterior pelvic ring injuries.

OBJECTIVETo separate and identify chemical constituents from stem barks of male plants of Populus tomentosa.

METHODFresh stem barks of P. tomentosa were extracted with methanol to obtain extracts which were suspended in water and blended successively with petroleum ether, ethyl acetate and n-butanol. Various chromatographic techniques were used to separate and purify the constituents extracted with ethyl acetate and n-butanol fractions. Their structures were identified on the basis of their physicochemical properties and spectral data.

RESULTTwelve compounds were separated with ethyl acetate and n-butanol fractions and identified as benzoic acid (1), daucosterol (2), tremuloidin (3), rhamnocitrin (4), sakuranetin (5), 7-O-methylaromadendrin (6), isograndidentatin A (7), siebolside B (8), sakuranin (9), micranthoside (10), alpha-D-glucopyranose (11), and sucrose (12).

CONCLUSIONCompounds 4-12 were separated from this plant for the first time. Of them, compound 10 was separated from this plant genus for the first time.

Flavonoids ; analysis ; Glucosides ; analysis ; Phenols ; analysis ; Populus ; chemistry

OBJECTIVETo research the effect of polysorbate 80 (Tween 80) on Yuxingcao injection and volatile oils from Houttuynia cordata.

METHOD1H-NMR spectra of aldehydic and new matter in Yuxingcao injection, volatile oils of H. cordata, and solutions of Tween 80 and volatile oil of H. cordata are determined and compared from various angles of growing origin, storage temperature, and storage time.

RESULTThree aldehydic singlets in 1H-NMR spectra of every volatile oil from 4 aerial part of H. cordata were observed. These aldehydic peaks were basically disappeared and a new peak at delta 8.30 was found in 1H-NMR spectra of the volatile oil solutions in tween 80. Any obvious aldehydic peak in 1H-NMR spectra did not be observed in Yuxincao injection. A weak peak at 8 8.30 was found in 1H-NMR spectra in Yuxincao injection, and the peak high of delta 8.30 was remarked gone up when the injection was stored in 40 degrees C for 1 to 3 months.

CONCLUSIONTween 80 might cause the obvious reduce of aldehydic compounds contents and the production of a novel singal at delta 8.30 in 1H-NMR spectra when it was mixed with the volatile oil from the aerial part of H. cordata. The novel signal at delta 8.30 in 1H-NMR spectra existed in Yuxincao injection and was very small, but was increased remarkably when the Yuxincao injection was stored at 40 degrees C for 1 month at least.

Chemistry, Pharmaceutical ; instrumentation ; Drugs, Chinese Herbal ; chemistry ; Houttuynia ; chemistry ; Oils, Volatile ; chemistry ; Plant Oils ; chemistry ; Polysorbates ; Temperature

Objective To investigate the role of neutrophils and their IgA Fc receptor CD89 in the occurrence of Henoch-Sch（o）nlein purpura (HSP),to evaluate their effects on vascular endothelial cell apoptosis,and to explore their mechanisms.Methods Peripheral blood neutrophils were isolated from 30 children with acute HSP and 9 age-matched healthy controls separately.After isolation of serum IgA by Jacalin affinity chromatography,IgA was purified by polypropylene dextran gel chromatography.Real-time fluorescence-based quantitative PCR (qPCR) and Western blot analysis were performed to determine the mRNA and protein expression of CD89 on neutrophils respectively,and flow cytometry was conducted to measure the expression of neutrophil activation marker CD11b.Human umbilical vein endothelial cells (HUVEC) were co-cultured with neutrophils isolated from patients with HSP (HSP group) and healthy controls (healthy control group) separately.Moreover,the HSP group were divided into 3 subgroups to be treated with serum IgA isolated from the HSP patients (HSP IgA group),monomeric IgA (mIgA group) and phosphate-buffered saline (blank control group) respectively.Then,flow cytometry was conducted to detect apoptosis of co-cultured HUVEC,and enzyme-linked immunosorbent assay (ELISA)to measure levels of interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-α) in the supernatant of co-cultured cells.Results There was no significant difference in the mRNA expression of CD89 on neutrophils between the patients with HSP and healthy controls (P =0.98),but the protein expression of CD89 was significantly lower in the patients with HSP than in the healthy controls (0.60 ± 0.16 vs.0.83 ± 0.24,P =0.03).The expression of CD1 1b on neutrophils was significantly higher in the patients with HSP than in the healthy controls (1 880.25 ± 388.29 vs.1 109.25 ± 364.25,P ＜ 0.01).The apoptosis rate of co-cultured HUVEC was also significantly higher in the HSP group than in the healthy control group (37.44％ ± 5.49％ vs.17.14％ ± 4.45％,P ＜ 0.01).In addition,the H SP IgA group showed significantly higher apoptosis rate of cocultured HUVEC and levels of IL-8 and TNF-cα in the supematant compared with the mIgA group (all P ＜0.01) and blank control group (P ＜ 0.01,=0.01,=0.02,respectively).Conclusions Peripheral blood neutrophils in patients with HSP are activated,which can induce the apoptosis of vascular endothelial cells.HSP IgA can promote the neutrophil-mediated apoptosis of vascular endothelial cells and secretion of IL-8 and TNF-α,while mIgA may show a certain inhibitory effects.