Objective To explore the expression levels of RAS-interacting protein 1 (RASIP1) mRNA and protein in hepatocellular carcinoma (HCC) tissues and its cell lines,and to analyze the relationship between RASIP1 and tumorigenesis and progression of hepatocellular carcinoma.Methods The expression levels of RASIP1 mRNA and protein in 29 hepatocellular carcinoma tissues and the corresponding adjacent non-cancer liver tissues (ANLTs),as well as those in the HCC cell lines such as LO2,HEPG2,MHCC97-H and HCCLM3 were detected using real-time PCR and western blot.Results The RASIP1 expression levels decreased significantly in HCC tissues when compared with the corresponding ANLTs; The expression levels of RASIP1 mRNA and protein in LO2 were significantly higher than those in other HCC cell lines (P ＜ 0.05) ; The expression levels of RASIP1 mRNA and protein in MHCC97-H and HCCLM3 were significantly lower than those in HepG2 (P ＜ 0.05).Conclusions HCC tissues had lower expression than those in ANLTs.On analyzing the RASIP1 levels of HCC tissues and its cell lines,we speculated that RASIP1 might suppress recurrence and metastasis of HCC.

The financial crisis has made the graduate of grade 2009 career have undergone profound changes in mentality,How to strengthen the ideological and political education of graduates and career guidance is a new task we have to face.In this paper,through a questionnaire survey,students’collective conversation,individual visits,the case summary,we understand the mental state of college graduates in 2009 session in Shenyan,and have analyzed it to provide a basis for career guidance for the targeted colleges and universities under the new circumstances.

Objective:To explore the risk factors of neurological complications after interventional treatment of ruptured intracranial aneurysms(RIAS), and to establish a predictive model of nomogram.Methods:The clinical data of 89 patients with RIAS who underwent endovascular treatment in Nanyang Second General Hospital Affiliated to Xingxiang Medical University from January 2016 to January 2019 were retrospectively studied.The clinical imaging data were collected and followed up for 6 months.The patients were divided into two groups: no neurological complications group (61 cases) and neurological complications group (28 cases). To analyze the clinical indicators and the possible related factors of neurological complications after RIAS interventional therapy.A nomogram was established to score the influencing factors, and a scoring prediction model was constructed; the clinical calibration of the model was evaluated by consistency index (C-index) and calibration curve, and the clinical differentiation of the model was evaluated by nomogram relying on ROC curve.Results:Multivariate logistic regression analysis showed that Hunt-Hess classification ( OR=4.927, 95% CI: 1.189-20.426, P=0.028), Fisher classification ( OR=4.633, 95% CI: 1.012-21.208, P=0.048 ), aneurysm cyst xiaofu ( OR=5.918, 95% CI: 1.104-24.948, P=0.015), wide carotid aneurysm ( OR=4.381, 95% CI: 1.029-18.645, P=0.046) and treatment Strategy ( OR=4.887, 95% CI: 1.235-19.329, P=0.024) is an independent risk factor for nerve-related complications after RIAs interventional therapy.The predictive model of nomogram showed that Hunt-Hess classification (grade IV, V) was 100, aneurysm bleb (with) 98, treatment strategy (stent implantation) 95, wide-necked aneurysm (yes) 92 and Fisher grade (grade III, IV) 81; the C-index of the predictive model was 0.871; the nomogram relied on ROC curve AUC 0.871, and the treatment strategy (stent implantation) was 95; the Fisher grade (grade III, IV) was 81; the C-index of the predictive model was 0.871.The sensitivity and specificity were 85.71%(24/28) and 77.05%(47/61) respectively. Conclusion:Hunt Hess classification, Fisher classification, aneurysmal sac caruncle, wide necked aneurysms and treatment strategies will affect the occurrence of neurological complications after RIAS interventional therapy.The nomogram established by this method can provide intuitive and reliable reference for clinical practice.

Objective To compare the effect of cimetidine associated with bletilla striata,rhubarb on peptic ulcer,cushingulcer,acute erosive gastritis,bleeding of esophagus and gastric funds varicosis of cirrhotic portal hyper- tension and gastri cancer.Methods 136 patients with upper gastrointestinal hemorrhage were randomly divided into 2 groups.The treated group(n=69)was given cimitidine associated with bletilla striata,rhubarb and the control group(n=67)was given cimitidine only.The effects of hemostasis in 2 groups were observed.Results The effect of hemostasis in the treated group was better than that in the control group on peptic ulcer,cushingulcer,acute ero- sive gastritis and gastric cancer(P＜0.05).There were no difference between 2 groups about bleeding of esophagus and gastric funds varicosis of cirrhotic portal hypertension(P＞0.05).Conclusion The effect of hemostasis on up- per gastrointestinal hemorrhage treated by integrated traditional Chinese medicine and western medicine might achieve better on cushingulcer,acute erosive gastritis,peptic ulcer and gastri cancer.It has less adverse effect and more safety.

Objective Via targeted inhibition of oncogene Bmi-1 expression by RNAi interfering technology in vitro, to observe its effect on the proliferation and cell cycle of gallbladder cancer cells. Methods Four miRNABmi-1 recombinant plasmids were constructed according to different Bmi-1 sites. RT-PCR and Western blot were used to mRNA and protein expression of Bmi-1 in gallbladder cancer cells were measured by RT-PCR and Western blot. mRNA and protein expression of Bmi-1 in gallbladder cancer cells. The most effective interfering plasmids in the miRNABmi-1 groups were transfected into GBC-SD cells. Cell proliferation and cell cycle were analyzed 48 h after transfection by BrdU and flow cytometry. Results Bmi-1mRNA expression in miRNAbmi1-1,-3 and-4 was significantly lower than the control group (P<0.05);and Bmi-1 protein expression in miRNAbmi1-2,-3 and-4 was significantly lower than the control group (P<0.05). The recombinant plasmid in miRNAbmi1-4, with the strongest inhibitive effect of Bmi-1mRNA and protein expression, was transfected into GBC-SD cells,then the cell proliferation rate (46.63 ± 5.31) was significantly lower in mRNABmi1-4 group than the control groups (P<0.05);G0/G1 phase cells increased (72.20 ± 1.71) and G2/M and S phase cells decreased (18.30 ± 7.21, 9.50 ± 6.01) in miRNABmi1-4 group. Both were significantly different from the control groups (P<0.05). Conclusions Targeting and silencing Bmi-1 expression can effectively inhibit the proliferation of GBC-SD cells and restrain the cell cycle atin G0/G1 phase. Bmi-1 gene may be a novel target for geneic therapy of gallbladder carcinoma.

Objective To observe the variations in intraocular pressure(IOP)in vocal cord polyp resections supported by pedestal Iaryngoscope with Tono-Pen tonometer.Methods The IOP of patients (grade Ⅰ-Ⅱby ASA)who underwent vocal cord polyp resections supported by pedestal laryngoscope were detected by Tono-Pen tonometer 5 minutes later on supine position before the operation(T1),5 minutes later on cervical hyperextension position before the operation(T2),5 minutes later on cervical hyperextension position after the operation(T3),5 minutes lateron supine position after the operation(T4),20 minutes later on supine position after the operation(T5)after general anesthesia respectively.At each point the changes of mean arterial pressure(MAP),heart mte(HR),end-tidal carbon dioxide partial pressure(PETCO2),and airway pressure(PAW)were observed as well.Results There were no differences in MAP,HR,RETCO2,PAW at each point statistically.The IOP increased significantly at T2,T3,T4 compared with IOP at T1[(19.0±1.8),(25.7±1.9),(17.8±1.9)mm Hg(1 mm Hg=0.133 kPa)vs(11.9±1.7)mm Hg](P＜0.05).The differences between IOP at T2 and T3 were manifest(P＜0.05).So it Was the situation when the IOP at T3 and T4,T4 and T5 were compared(P＜0.05).The IOP at T5 was(12.1±1.5)mm Hg,there was no difference compared with T1.Conclusion The IOP increases gradually from the point when the patient put on cervical hyperextension position before the operation after general anesthesia and achieves the summit when the patient put on cervical hyperextension position after the operation,finally,decreases back to the preoperative level when the patient put on supine position after the operation.

BACKGROUND: Studies demonstrated that small intestinal submucosa (SIS) had no immunogenicity, which can not lead to rejection following transplantation, thus, this is an ideal skin substitutes for natural skin.OBJECTIVE: Basic fibroblast growth factor (bFGF) gene was transfected into bone marrow mesenchymal stem cells (BMSCs),and combined the cells with SIS to construct tissue-engineered skin.METHODS: BMSCs were obtained from Japanese big-earad rabbits, and in vitro cultured. Then the subculturad BMSCs were transfected by pCDNA3.1 plasmid, followed by incubation on swine SIS to construct the tissue-engineered skin. The growth of cells and phenotype of BMSCs were detected by flow cytometry. In addition, the result of transfecting BMSCs with pCDNA-bFGF vector was measured by Western blot, and the structure of tissue-engineered skin was observed.RESULTS AND CONCLUSION: After passaged, BMSCs were grown quickly with Iong-fusiform shape. The cells were positive expressed CD90 and CD44, but negative expressed CD45. bFGF had been transfected into BMSCs, and stable expressed. The transfected BMSCs grew well in SIS. By this method, tissue-engineered skin can be constructed in vitro.

OBJECTIVE: To investigate the in vivo possibility of osteogenesis and angiogenesis of tissue-engineered periosteum in rabbits.METHODS: The marrow mesenchymal stem cells (MSCs) derived from New Zealand rabbits were adhered to small intestinal submucosa (SIS) to fabricate the tissue-engineered periosteum. Totally 12 New Zealand rabbits were received critical bone defect in bilateral radii to prepare models. The tissue-engineered periosteum was randomly implanted in one side of bone defect,and the other side was treated by SIS. At 4 weeks after operation, the angiogenesis of tissue engineered bone was detected by Tetracycline fluorescence microscopy and formaldehyde-ink perfusion method; simultaneously, the new bone formation was firmed by haematoxylin-eosin staining.RESULTS: Animals showed normal daily behaviors and non-infection wounds healing. The gross observation showed that bone defects in the experimental side were bridged with newly formed bone; while the defects of the control side were remained empty.Tetracycline fluorescence microscopy and hisotological examination could confirm the new bone tissue formation in the experimental side. The ink staining in new bone specimens suggested that there were abundant of neovasculization in tissue-engineered bone.CONCLUSION: Tissue-engineered periosteum can form new bone in allogenic rabbits and can be vascularized by some inherent mechanism for new bone tissue survivor.

BACKGROUND: Basic fibroblast growth factor (bFGF) has significant promotion effects on repair in trauma, but local application cannot play a role for a long time.OBJECTIVE: To observe expression of bFGF gene in rabbit bone marrow mesenchymai stem cells (BMSCs) following transfection.DESIGN, TIME AND SETTING: The cytogene in vitro observation was performed at the College of Life Science, Yunnan University from March to August 2009.MATERIALS: One Japan flap-eared rabbit was purchased from the Department of Animal, Kunming Medical College. pCDNA3.1plasmid (Invitrogen, USA) was used in this study.METHODS: Bone marrow was extracted from rabbit anterior superior iliac spine. BMSCs were harvested by the adherent method.Cells were digested and subcultured when 80% confluent. According to GeneBank bFGF cDNA sequence, gene was designed and synthesized. Following electropherosis, the gel was retrieved using xhol I, BamH I enzyme digestion. Restriction enzyme was used to perform enzyme digestion, electropherosis and gene recovery in PcDNA Vector plasmid. bFGF DNA was connected with PcDNA Vector plasmid. PcDNA-bFGF eukaryotic expression vector was constructed. Recombinant was transfected into rabbit BMSCs using liposome infection protocol, and stable transfected line was screened.MAIN OUTCOME MEASURES: BMSC surface antigen expression was measured. Western blot was utilized to determine the expression of target protein.RESULTS: Results of flow cytometry showed that cultured cells were positive for CD90 and CD44, but negative for CD45.Results of immunohistochemistry demonstrated that vessel-derived BMSCs were negative for CD34, but positive for CD44. In cell disruption liquid of bFGF-transfected BMSCs, a significant positive zone of hybridization was visible at M, 23 000. However, no positive band was found in protein from pCDNA3.1(-) blank vector-transfected BMSCs.CONCLUSION: The bFGF gene was successfully transfected into BMSCs, and this target gene can stably express.

OBJECTIVETo observe mainfestations of syndrome and biochemical indices of hypertensive model rats with excessive accumulation of phlegm-dampness syndrome (EAPDS), and to explore its possible pathological mechanism.

METHODSEAPDS rat model was prepared in 50 Wistar rats by feeding with high fat forage. Meanwhile, a normal control group consisting of 10 Wistar rats was set up by feeding with normal forage. After 25-week continuous feeding, 22 rats with body weight (BW) and blood pressure (BP) exceeding 25% those of the control group were selected as a model group. BW, BP, blood lipids, and related serological indicators were detected in all rats. Morphological changes of target organs were observed. mRNA expression levels of leptin receptor (LepR), Janus kinase2 (Jak2), signal transducer and activator of transcription 3 (Stat3), suppressor of cytokine signaling-3 (Socs3), angiotensin II receptor type 1 (AT1), angiotensin II receptor type 2 (AT2), phosphatidylinositol 3 kinase (P13K), serine threonine kinase (Akt), nuclear factor of kappa B (NF-κBp65), inhibitor of nuclear factor kappa-B kinase α (IKKα), NF-kappa-B inhibitor β (lKKβ), NF-kappa-B inhibitor α (IKBα), and AMP-activated protein kinase (AMPK) were detected by quantitative real-time PCR (qPCR). Expression levels of AT1 and LepR in aorta were detected by immunohistochemical assay and Western blot respectively.

RESULTSCompared with the control group, BW, BP, and blood lipids increased; serum levels of leptin (Lep) , Ang II, Hcy, ET-1, TNF-α, IL-6, and p2-MG increased, but NO decreased in the model group (P < 0.05, P < 0.01). Aortal endothelial injury and smooth muscle cell proliferation occurred in the model group, accompanied with heart and renal injury. Compared with the control group, mRNA expression levels of LepR, Jak2, Stat3, Socs3, AT1 , PI3K, Akt, NF-κB p65, IKKβ, IKBα, and AMPK in aorta were up-regulated significantly (P < 0.05), while the expression of IKKa decreased (P < 0.05). Immunohistochem- ical staining showed, brownish yellow deposit of AT1 and LepR was obviously increased, with more extensively positive distribution. Western blot results showed, as compared with the control group, protein expression levels of AT1 and LepR obviously increased in the model group (P < 0.05).

CONCLUSIONSModel rats exhibited typical syndromes of EAPDS. They put up weight with fat abdomen, gloomy hair, poor appetite, hypersomnia, lowered activities , reduced food intake, loose stool, dark red tongue, white tongue with white, thick, greasy fur. Lep could be taken as one of objective indicators for evaluating hypertension rat model with EAPDS.

Animals ; Aorta ; Cell Proliferation ; Disease Models, Animal ; Hypertension ; physiopathology ; I-kappa B Proteins ; Interleukin-6 ; Leptin ; blood ; NF-KappaB Inhibitor alpha ; NF-kappa B ; Phosphatidylinositol 3-Kinases ; Rats ; Rats, Wistar ; Suppressor of Cytokine Signaling Proteins ; Transcription Factor RelA ; Tumor Necrosis Factor-alpha