Objective To construct expression vectors of human ribosomal protein S 3(RPS3) and RPS3Ser209 mutant in orcler to investigate the effect of RPS3Ser209 mutant on NF-κB signaling pathway and DNA binding capacity .Methods The vector RPS3-myc was amplified by polymerase chain reaction ( PCR) from the human liver cDNA and subcloned into pcDNA-3.1myc-HisB.RPS3S209A represented mutant RPS3 expression vectors, in which the designated amino acid was mutated to an alanine residue .Dual luciferase reporter gene assay was used to detect the NF-κB transcription activity in HEK293 cells,immunofluorescence to detect RPS3 location, and EMSA to examine NF-κB DNA-binding activity.Results The expression vectors of RPS3-myc and RPS3S209A-myc were constructed.Compared with wild-type RPS3,the nucleus translocation, transactivation activity of NF-κB and DNA binding ability of RPS3S290A were reduced significantly .Conclu-sion The impact of RPS3 on NF-κB signaling pathway depend on its serine 209.

BACKGROUND: Type Ⅰ collagen is a specific collagen secreted by in vitro cultured osteoblast, and the formed network is the basis of bone mineralization, which also reflects the ability of osteoblast bone formation. Studies have shown astragalus root increased osteoblast proliferation. However, the effect of astragalus root on improving type Ⅰ collagen expression of osteoblast remains poorly understood.OBJECTIVE: To evaluate the effect of astragalus root injection on the abilities of rat cranium-derived osteoblast proliferation and type Ⅰ collagen expression.METHODS: Rat osteoblast was cultured in vitro and divided into control group (MEM culture solution containing calf serum) and astragalus root groups (different concentrations). The effect on osteoblast proliferation was evaluated on days 1, 3, 5, 7, and 9 by MTT method. Moreover, the expression of type Ⅰ collagen protein was observed after 6 hours of treatment with astragalus root injection using in cell western-blot method. In addition, the gene expression of COLLal was investigated by real-time PCR method.RESULTS AND CONCLUSION: From days 3 to 9, the different concentrations of astragalus root injection improved osteoblast proliferation, respectively compared with control group (P ＜ 0.05), and this ascending trend peaked on day 7. Different concentretions of astragalus root injection improved COLLol mRNA expression, especially 15% astragalus root injection was the most effective. The type Ⅰ collagen protein expression of 15% and 10% astragalus root injection were significantly greater compared with the control group (P ＜ 0.05). Astragalus root injection improved in vitro cultured osteoblast proliferation and type Ⅰ collagen secretion in a certain dose-effect manner.

BACKGROUND:In 1990s,overseas researchers use balloon occlusion method for establishing closed-chest animal models of myocardial infarction. But,ventricular fibrillation and thrombosis of intraoperative factors reduce the success rate of establishing the models. Currently,there are a few reports on establishing the large animal models. OBJECTIVE:We used balloon occlusion method for establishing closed-chest swine models of myocardial infarction,and explored ways to improve the success rate of modeling. DESIGN,TIME AND SETTING:The randomized controlled animal study of pathology observation was performed at the Department of Cardiology,First Affiliated Hospital of Kunming Medical College and Research Room of Pathology,Kunming Medical College from July 2008 to May 2009. MATERIALS:Fifteen Diannan small-ear pigs weighing 19-25 kg,aged 8-11 months,were divided into three groups:sham operation group,ischemia-reperfusion group,and ischemic postconditioning group,with 5 pigs in each group.METHODS:After the coronary occlusion and reperfusion period,the prophylactic use of lidocaine (1.0-2.0 mg/kg) infusion to control arrhythmia,and use of heparin to prevent and treat the thrombosis. A balloon catheter was positioned in the distal end of the first diagonal branch of the left anterior descending (LAD) artery under fluoroscopic guidance. In the sham operation group,the balloon was only placed to the LAD,did not block coronary artery. In the ischemia-reperfusion group,inflatable balloon occlusion was done for 60 minutes in the LAD after the balloon removed. In ischemic postconditioning group,after the balloon was inflated and occluded the LAD for 60 minutes,ischemic postconditioning was elicited by eight cycles of 30-second reperfusion,followed by 30-second reocclusion.MAIN OUTCOME MEASURES:Coronary angiography,electrocardiogram (ECG) and cardiac enzymes test was conducted to evaluate models of myocardial infarction. After three days,cardiac 2,3,5-triphenyltetrazolium chloride (TTC) staining and pathological examination was done to verily myocardial infarction.RESULTS:In the sham operation group,all pigs survived. In the ischemia-reperfusion group,4 pig models of myocardial infarction were successfully established,and one died of refractory ventricular fibrillation. In the ischemic postconditioning group,models of myocardial infarction after ischemia were successfully established. Following distal left anterior descending artery occlusion,the ECG leads V13 on the ST-segment elevation,the sick rational Q-wave formed;myocardial enzyme evolution of myocardial infarction in the human body was basically the same process. The site of myocardial infarction,basically the same parts,was located in apical,left ventricular anterior wall,and the former interval. TTC staining was normal myocardium brick red,myocardial infarct area appeared pale;pathological examination revealed a normal structure of myocardial infarct damage,cytoplasm condensed,dyeing deepening,transverse striations disappeared,nuclear enrichment,dissolution,fragmentation,many erythrocytes around the infarct area with abundant granulation tissue and a large infiltration of inflammatory cells.CONCLUSION:The described model presents a less invasion to the animals,and is the closest to the process of clinical practice.Intraoperative use of lidocaine and heparin for controlling arrhythmia and thrombosis of the models is successfully established as an effective manner. Ischemic postconditioning may be one of the factors for improving the modeling success rate.

Lesch-Nyhan syndrome(LNS) is a congenital X-linked recessive inherited disorder caused by mutations in the hypoxanthine guanine phosphoribosyl transferase (HPRT) gene.A deficiency of the HPRT enzyme is responsible for the disease.The main clinical manifestation includes hyperuricemia, juvenile-onset gouty arthritis and neurological developmental disorders.Studies have reported there are more than 400 HPRT gene mutation sites, but the incidence of LNS in the Chinese population is extremely low.Here we report a 16-year-old male patient who suffered neurological dysfunction at an early age and gouty arthritis in his youth.DNA of patient and his family members were extracted from peripheral blood lymphocytes.The coding region and the intron-exon boundaries of HPRT gene were sequenced by standard methods.We found a mutation in exon 3 of the HPRT gene of the patient and his mother (Exon3:c.143G>A), which resulted in an arginine to histidine (p.R48H) substitution in the encoded protein.No activity of the enzyme HPRT was detected in the erythrocytes.The same mutation was reported in several European families, but was found in Chinese family for the first time.Clinicians in China have poor experience in diagnosing LNS case, due to the low incidence in China.Therefore LNS screening for infants or adolescents with hyperuricemia, gouty arthritis and neurological dysfunction should be performed.

Objective To investigate the relationship of antithrombin‐Ⅲ activity and thrombosis risk in liver cirrhosis with Child‐Pugh classification .Methods In our hospital from June to December 2014 ,60 liver cirrhosis patients were selected randomly included into this experiment group ,The 60 cases of control group were from medical examination of health in our hospital .The plasma AT‐Ⅲ activity and D‐D concentration in all these cases were detected and analyzed .Results The AT‐Ⅲ in cirrhosis patients were significantly lower than which in healthy persons(P<0 .05) .The lower level of AT‐Ⅲ is in these patients which were in seri‐ous condition(P<0 .05) ,the abnormal rate of D‐D concentration is also higher at the same time(P<0 .05) .Conclusion The detec‐tion of AT‐Ⅲ level in patients with liver cirrhosis is directly related to the severity of clinical and thrombosis risk .The AT‐Ⅲ de‐tection level can be used to judge the patient′s condition and develop appropriate treatment strategies .

Objective To summary and discuss the curative effect and experience of repairing serious and complicated soft tissue defects of traumatic shank by flap or musculocutaneous flap by anatomosis.MethodsFrom October 2009 to December 2011, the wounds of 59 patients suffering from serious and complicated soft tissue defects of traumatic shank were covered by VSD after repeated debridement,when the conditions of the whole bodies were stable and the local acute infection was controlled in the main. Fifty-six patients were repaired by a single flap or musculocutaneous flap attributing to the fresh granulation, three patients were repaired by compound tissue flaps because a single flap was insufficient. Five patients who had no available blood vessels at recipient site were repaired by flaps with bridge cross vascular anatomosis. The compound flaps were no more than 2 pieces, the maximum area of flap was 32 cm × 13 cm, the minimum was 15 cm × 8 cm.Results The flaps of all of 59 cases survived, fifty-seven cases were healed in one stage, two in two stage, one case had complication of infection at donate site due to the hematocele, and was cured by debridement and skin graft. Serious and complicated soft tissue defects of shanks were repaired by reconstruction,damaged limbs were salvaged,the functions of the legs were reserved. Conclusion It is the most effective and irreplaceable way that using flaps and musculocutaneous flaps,by anatomosis with the microsurgery technique, repairs the serious and complicated soft tissue defects of shank, which can shorten course and salvage the damaged limbs.

Objective To investigate the protective effects of atorvastatin on hyperphosphate-induced rat vascular smooth muscle ceils (RVSMCs) calcification and to discuss the mechanism. Methods RVSMCs were placed in various culture media, including normal phosphate medium, high phosphate medium, ZVAD-FMK medium and atorvastatin medium.Calcium content and cell protein content were quantified by the o-cresolphthalein complexone method and BCA protein assay respectively. Calcification was visualized by yon Kossa staining. And cell apoptosis was quantified by ELISA. Results (1)At day 3, 6, 9, RVSMCs calcification occurred more frequently in high phosphate medium than that in normal phosphate medium (P<0.05). (2)At day 6, RVSMCs calcification was significantly inhibited in 1.0 μmol/L and 2.0 μmol/LZVAD-FMK medium (P<0.05). And in 10 nmol/L and I00 nmol/L statin medium, RVSMCscalcium deposition significantly decreased (P<0.05). (3)RVSMCs apoptosis and calcification occurredfrequently in high phosphate medium. And atorvastatin significantly inhibited RVSMCs apoptosisboth in long-term and short-term (P<0.05). Conclusions Hyperphosphate can induce the calcium deposition of RVSMCs in vitro. Atorvastatin protects RVSMCs from phosphate-induced calcification by inhibiting apoptosis.

Objective To assess the long efficacy and safety of Lamotrigine(LTG) monotherapy and add-on therapy different types of epileptic seizures in children.Methods According to the classification of the 1981 and 1989 International Union of Antiepileptic for epileptic seizure,a total of 124 cases with epilepsy were included in the study and divided into non-refractory group with 93 cases and refractory group with 31 cases.LTG treatment only or add-on were used.Original drug dosage was not changed and LTG was added slowly and carefully untill the terrible side effect appeared.The average monthly seizure frequency with baseline in the last 3 months was compared and the side effect was observed.Results Total efficiency was 72.6%,control rate was 51.6%.Total efficiency and control rate of the non-refractory group was 81.0% and 61.3%,which was significantly higher than those of refractory group(48.4%,22.6%).Total efficiency and control rate of the combination group with LTG and valproate sodium(VPA) was 78.4% and 54.5%,which was significantly higher than those of the group of LTG only(61.0%,44.0%).Clinical results was different significantly between the course of the observation period within and over 5 years(P

Objective To investigate the relationship between high-sensitivity C-reactive protein(hsCRP), pulse pressure(PP) and microalbuminuria(MAU) in patients with essential hypertension.Methods Consecutive 60 patients with essential hypertension were classified based on the MAU level:patients with MAU(n=30) and the group of patients with normoalbuminuria(NMAU,n=30) with 30 healthy subjects as control.MAU,hsCRP,PP were determined.Results The level of hsCRP and pulse pressure in MAU group were higher than those in NMAU patients and healthy subjects (P

Objective To evaluate the application of non-bioartificial liver support system (ALSS) combined with liver transplantation(LT) in treating mid-or end-stage chronic severe hepati- tis.Methods ALSS plus liver transplantation were employed in treating 28 patients with mid-or end- stage chronic severe hepatitis.Clinical data from the patients before and after treatment were collect- ed.The survive rate of ALSS plus LT group were compared with that of medication group 99 cases and medication plus ALSS group 30 cases.The data were analyzed with t test and X~2 test.Results After 57 times ALSS treatment,the serum total bilirubin(TBil),prothromin time(PT),bile acid, blood urea nitrogen(BUN),creatnine(Cr) and ammonia of all the 28 patients got improved(P 0.05).The clinical symptoms and signs of the patients were ameliorated at median 3 d(1～153 d). All patients were bridged to liver transplantation successfully after median 20 d(1～153 d).The 3 and 6 months post-operation survival rate of ALSS plus LT group(71.4%,71.4%) were significantly higher than those in medication group(18.2%,11.1%) and medication plus ALSS group(36.7%, 26.6%)(P