Objective To investigate the effect of tumor necrosis factor-α(TNF-α)on cytokines expression in mice with different tidal volume(VT). Methods Thirty six SPF male BALB/c mice were randomly divided into three groups:A group(spontaneously breathed 4 h),S and L group(mechanically ventilated 4 h,VT=6 mL/kg and VT=20 mL/kg). The three groups were then randomly divided into two subgroups(C and T group,n=6). Mice in C group were injected with NS,while that of T groups were injected with TNF-α at 20 μg/kg from tail intravenous. Morphological manifestation was observed by hematoxylin and eosin staining. Ultrastructure of macrophages were examined by transmission electron microscopy. The expression of TNF-α,interleukin(IL)-1β,IL-6,IL-10 in serum and BALF were measured by ELISA. The expression of NF-κB mRNA and protein were detected by RT-PCR and WB,respectively. Results Compared with A and S group,following could be observed under light microscope in L group:the pulmonary interstitial edema,consolidation of part of the lung tissue,alveolar space fusion and a number of inflammatory cells infiltration. The expression of TNF-α,IL-1β,IL-6,IL-10 in serum and BALF,the expression of NF-κB mRNA and protein in macrophages of L group were significantly higher than those of A and S group(both P<0.05). The expression of these cytokines and NF-κB mRNA and protein were increased in T group (P < 0.05). The expression NF-κB is positively correlated with the expression of TNF-α,IL-1β and IL-6. Conclusion TNF-α is a key factor in inflammation of ventilation induced lung injury,which may provide a new method for prevention of VILI in clinical practice.

Objective Evaluating the effect of situational questions for qualifying the students'abilities in physiolo-gy examination .Methods Comparing the difficulty coefficients and discrimination indexes between situational and traditional choice questions .Results Compared with the traditional choice questions , the difficulty coefficient of situational questions increased , while the discrimination indexes were more reasonable .The discrimination indexes of situational understanding questions were higher than those of the traditional memory and understanding questions . There were no difference between discrimination indexes of the situational application questions and those of tradi -tional application questions .Conclusions Situational questions not only improved the quality of examinations , but also facilitate evaluating students'learning ability .

Objective To evaluate the value of combined hepatectomy in radical resection for hilar cholangiocarinoma. Methods The clinical data and follow-up data of 67 patients of resection for hilar cholangiocarinoma in Henan Tumor Hospital from June 2005 to october 2008 were retrospectively analyzed. Results According to intraoperative exploration situation and bismuth types, tumor resection was combined performed with hepatectomy (n＝38)or non-hepatectomy (n＝29). The rate of R0 resection was 55.3% in hepatectomy group(n＝21) and 34.5% in non-hepatectomy group(n＝10), and the difference was significant（P＝0.024）. The incidence of complications were 39.5% in hepatectomy group(n＝15) and 13.4% in non-hepatectomy group(n＝4), and one patient with liver and kidney failure died in hospital. The 1, 3, 5 years of survival rate were 89.3%,53.6% and 32.1% respectively in R0 group (n＝31) and 69.7%,30% and 10% respectively in R1～R2 group(n＝36), there were significant differences in the postoperative survival rate between both groups（P＝0.018). The 1, 3, 5 years of survival rate were 81.8%,48.5% and 24.2% in hepatectomy group and 75%,32% and 16% in non-hepatectomy group respectively, and the differences were significant（P＝0.037). Conclusions Aggressive resection including combined hepatectomy for hilar cholangiocarcinoma can play an important role for curative effect and long term survival rate.

OBJECTIVEIn order to identify a species of Uncaria, molecular phylogenetic analysis was carried out by using the rDNA ITS sequence as molecular marker.

METHODTotal DNA was extracted from the plant with modified CTAB method and thereby rDNA ITS regions were amplified with universally conserved primer. The rDNA ITS amplicon was characterized by cloning, sequencing, blasting in GenBank and phylogenetic analyses using PAUP by maximum parsimony (MP) criteria.

RESULTThe rDNA ITS entire sequence of this species of Uncaria was 719 bp. The sequence is related to the U. sinensis available in GenBank and the similarity reaches 99.7%.

CONCLUSIONBased on molecular biology methods of rDNA ITS region analysis, molecular identification is available in accurate classification on this species of Uncaria.

DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Phylogeny ; Sequence Homology, Nucleic Acid ; Species Specificity ; Uncaria ; classification ; genetics

OBJECTIVE:To reduce the incidence of irrational medical orders for parenteral nutrition,and promote the rational use of parenteral nutrition. METHODS:The medical orders for parenteral nutrition of the first quarter of 2016 in general surgery de-partment of our hospital were collected,and the number and types of its irrational medical orders were summed up. Then FO-CUS-PDCA(Find-organize-clarify-understand-select-plan-do-check-act)cycle management was adopted to analyze and improve the existing problems in issuing medical orders for parenteral nutrition. The improved(the third quarter of 2016)medical orders for par-enteral nutrition were collected,the number and types of its irrational medical orders were summed up,and management effect was evaluated. RESULTS:Establishing nutrition support group,strengthening the training and communication of medical staff,adding prescription evaluation module for parenteral nutrition in hospital information system and a number of measures had made the inci-dence of irrational medical orders for parenteral nutrition in general surgery department declined from 48.25%(1433/2970)before improvement to 5.67%(120/2118)after improvement. The incidences of cation excess,inappropriate selection of drugs and inap-propriate compatibility in irrational types were 0. CONCLUSIONS:FOCUS-PDCA cycle management can reduce the irrational rate of medical orders for parenteral nutrition and promote the rational use of parenteral nutrition in hospital.

Objective To investigate the associations of single nucleotide polymorphism of ( SNP ) of rs77418916 and rs8108402 in miR-181 with the risk of systemic lupus erythematosus ( SLE) in the Chinese population of Guangxi. Methods The lymphocyte sub-sets were analyzed by flow cytometry. The SNPs of miR-181 gene were detected by single nucleotide primer extension assay with SNaP-shot and DNA sequencing method. The relative expressions of miR-181a and miR-181c in mononuclear cells were detected by real time RT-quantitative PCR. Results The polymorphism of rs8108402 locus contained CC, CT and TT genotypes. The frequencies of CT and TT genotypes as well as the dominant and recessive model were different significantly between SLE group and control group ( CT vs CC:OR=1.50, 95％CI:1.03 to 2.19, P=0.033; TT vs CC: OR=2.65, 95％CI: 1.18 to 5.98, P=0.019; CC/CT vs TT: OR=2.23, 95％CI:1.01 to 4.93, P=0.048;TT/CT vs CC:OR=1.61, 95％CI:1.12 to 2.31, P=0.010) . The polymorphism of rs77418916 locus contain AA, AT and TT genotypes, but no association between rs77418916 polymorphism and susceptibility of SLE was found. The rel-ative expressions of miR-181a and miR-181c genes in SLE group were down-regulated compared with control group ( Z=-3. 22, P<0.01 and Z=-3.24, P<0.01, respectively) , and the patients carrying rs8108402 CT and TT genotype showed lower level of miR-181c compared with the patients carrying CC genotype (Z=-2.51, P<0.05). The absolute numbers of CD3+, CD4+, CD8+ and NK cells were decreased significantly in SLE group compared with that of control group ( P<0.01) . Conclusion The polymorphism of miR-181c rs8108402 may associate with the susceptibility of Chinese SLE patients in Guangxi region. The risk of SLE may increase in the individ-uals caring CT or TT genotype by decreasing the expression of miR-181c gene.

Objective:To establish the fingerprints of Microctis Folium by ultra high performance liquid chromatography (UPLC); To determine the contents of three flavonoids in the Microctis Folium; To evaluate the quality difference of Microctis Folium from different producing areas. Methods:The fingerprints were performed on Agilent ZORBAX SB C18 column (2.1 mm×150 mm,1.8 μm). The mobile phase was acetonitrile - 0.1 % acetic acid solution with gradient elution at a flow rate of 0.30 ml/min. The column temperature was 30 ℃ and the detection wavelength was 315 nm. The common fingerprint peaks were identified by UPLC-mass spectrometry, and the identification results were confirmed by comparison of reference materials. Waters Cortecs T3 C18 chromatographic column (2.1 mm × 100 mm,1.6 μm) was used for content determination. The mobile phase was methanol-0.1 % formic acid solution with gradient elution at a flow rate of 0.35 ml/min. The column temperature was 30 ℃ and the detection wavelength was 339 nm. The contents of vitexin, isovitexin and narcissoside in 15 batches of Microctis Folium from different habitats were determine. Results:There were 11 common peaks in the fingerprint of Microctis Folium. Identified by mass spectrometry and confirmed by reference substance,10 chemical components were identified, including caffeic acid, p-hydroxycinnamic acid, ferulic acid, vitexin, isovitexin, kaempferol-3-O-rutoside, astragaloside, narcissoside, isorhamnetin-3-O-glucoside and linden glycoside. The similarity between the fingerprints of 15 batches of Microctis Folium and the control fingerprint was greater than 0.95, indicating that the overall similarity of the fingerprints of Microctis Folium from different producing areas was high. The total contents of three active components were 3.23-5.64 mg/g in Yangjiang City, Guangdong, 3.60-5.78 mg/g in Zhanjiang City, Guangdong, 4.68-5.73 mg/g in Yulin City, Guangxi and 3.87-5.21 mg/g in Wuzhishan City, Hainan . There was no significant difference in the content of three active components in different producing areas. Conclusion:The fingerprints and the determination method established in the study are stable and feasible, which can be used for the quality evaluation of Microctis Folium.

Objective To evaluate the efficacy of the ratio of the fetal cardiac diameter to biparietal diameter( CBR) as a predictor of homozygous α-thalassemia-1 . Methods Single mid-pregnancies ( 15-22weeks) at risk of homozygous α-thalassemia-1 were enrolled . A total of 251 singleton pregnancies were recruited ,in which 63 cases were homozygous α-thalassemia-1 fetuses and the rest were unaffected . The CBR and cardiothoracic ratio(CTR) were measured by two-dimensional ultrasound . Then the accuracy of these variables were analyzed and compared with each other by ROC curves . Results ①The CBR and CTR in affected fetuses were significantly higher than those in the unaffected( P ＜0 .01) . ②With CBR＞0 .43 and CTR ＞ 0 .52 as the best cut off values ,the sensitivity and specificity of predicting homozygous α-thalassemia-1 fetuses in 15-22 gestational weeks were 95 .74% , 92 .06% and 94 .15% , 85 .71% , respectively ;the area under ROC curve were compared with Z test and there was no significant difference between them ( Z ＝ 1 .500 , P ＝ 0 .1335) . ③ When CBR and CTR were combined ,the sensitivity and specificity of the prediction were significantly increased ( the sensitivity of series experiment : 99 .75% ,the specificity of parallel experiment : 98 .87% ) . Conclusions CBR is a novel , effective and noninvasive predictor of homozygous α-thalassemia-1 in mid-pregnancy whose prediction efficiency is the same as traditional CTR . The measurement of CBR is easier to standardize and is not affected by thoracic lesions such as pleural cavity ,pericardial effusion and skeletal dysplasia . If combined with CTR ,it may play an important role in improving the prenatal detection rate of homozygous α-thalassemia-1 fetuses .

Hepatocellular carcinoma is a common malignant disease in clinical practice, and portal vein tumor thrombosis (PVTT) is one of the important factors affecting the prognosis of hepatocellular carcinoma. PVTT has strong oncologic characteristics and is highly susceptible to extrahepatic metastasis, complicating portal hypertension, leading to gastrointestinal bleeding or liver failure and causing death. In this paper, we review the formation mechanism of hepatocellular carcinoma combined with PVTT in terms of local anatomy, hemodynamics, molecular biology and tumor microenvironment to provide effective reference for clinical treatment.

This study aims to develop an improved cell screening system for farnesoid X receptor (FXR) agonists based on a dual luciferase reporter gene system. FXR response element (FXRE) fragments from FXR target genes were cloned and inserted into upstream of firefly luciferase (Luc) gene in the plasmid pGL4-luc2P-Hygro. In combination with the internal reference plasmid containing renilla luciferase, a dual luciferase reporter gene system was developed and used for high throughput screening of FXR agonists. After studying the effects of over-expression of RXR, mouse or human FXR, various FXRE fragments, and different ratio of FXR plasmid amount to reporter gene plasmid, induction efficiency of the screening system was optimized by the known FXR agonist GW4064, and Z factor for the system reached 0.83 under optimized conditions. In summary, an improved cell screening system based on double luciferase reporter gene detection system was developed to facilitate the discovery of FXR agonists, where a new enhanced FXRE element was formed by a superposition of multiple FXRE fragments from FXR target genes, instead of a superposition of traditional IR-1 (inverted repeats-1) fragments.
Humans ; Mice ; Animals ; Transcription Factors/genetics* ; DNA-Binding Proteins/genetics* ; Receptors, Cytoplasmic and Nuclear/genetics* ; Genes, Reporter ; Luciferases/genetics*