Objective To explore the adeB gene expression level impact of tigecycline for multidrug resistance Acinetobacter baumannii.Methods 57 strains multidrug resistance Acinetobacter baumannii were separated from September.2012 to September.2014.Epsilome test method was used to detect minimum inhibitory concentration of tigecycline against multi drug resistant Acinetobacter baumannii.PCR amplification detected adeB, adeR, adeS efflux pump gene distribution.Real-time fluorescent RT-PCR detection technology were used to detect relative expression level of each gene. Results 57 strains of multidrug-resistant Acinetobacter Bauman MIC value were 0.1-8μg/mL.The calculation of MIC50 =2μg/mL, MIC90 =4μg/mL. 49 strains sensitivity accounted for 86.0%, 5 intermediate strains accounted for 8.8%, 3 resistant strains accounted for 5.2%.57 strains of bacteria were divided into 38 strains lower sensitivity group and 19 strains sensitive group, with reduced susceptibility in lower sensitivity group 30 strains, accounting for 78.9%, sensitive group were all sensitive to tigecycline, accounting for 100%, for tigecycline susceptibility sensitive group was significantly higher than that of lower sensitivity group ( P <0.65 ) .41 strains were detected adeB, adeR, adeS gene in all strains, accounted for 86.0%, 29 strains were detected adeB, adeR, adeS genes in lower sensitivity group, accounted for 70.7%,12 strains were detected in sensitive group accounted for 63.2%, the difference between the two groups had no statistical significance.Fluorescent RT-PCR detection results showed that the relative expression of adeB gene with lower susceptibility group was significantly lower than that of sensitive group (P<0.05), relative expression levels of adeR and adeS gene in two groups had no significant difference.Conclusion AdeB gene plays an important role in tigecycline for multidrug resistance Acinetobacter baumannii, but may also exist outside of the adeB gene is involved in the regulation of resistance to tigecycline.

OBJECTIVE: To investigate the clinical application of tissue engineering for treating tendon injury.METHODS: A computer-based online search was conducted in Medline and China Academic Journals Database with the key words of "Biological material, Sinew mechanism heals" in both English and Chinese from 1974 to 2009. Relevant books were hand-retrieved.RESULTS: Scholars at home and abroad tried to investigate how to regulate cell proliferation, differentiation, and matrix synthesis by endogenous and exogenous growth factors so as to promote tendon repairing, relieve tendon adhesion, and decrease attenuation of biomechanical intensity. Tendon detect was mainly treated by autologous tendon transplantation,allogeneic tendon transplantation, artificial tendon transplantation, and tissue-engineered tendon transplantation. All those methods had both advantages and disadvantages. However, artificial biomaterial tendon has attracted much attention and it has been used for clinical application. Of course, there are still more problems to be solved, for example, source of tissue-engineered seed cells, immunological reaction of allogeneic tendon cells, specific mechanical intension, preparation and selection of degradable scaffold materials, correlation between cell and scaffold materials, and correlation between cell-material compound and peripheral tissues. With the development of tendon repairing and healing, single drug which was used to prevent tendon adhesion has developed into drug-barrier compound. Additionally, with the development of Chinese herb and molecular biological products, they not only inhibited exogenous healing, but also promote endogenous healing. Meanwhile, simple surgery has also developed into combined therapy.CONCLUSION: Although biomaterials for tendon repairing have achieved breakthrough in some respects, the results need also to be further studied.

There are two kinds of treatments on Alzheimer's disease (AD) rat by transplanting neural stem cells (NSCs),i.e.the replacing cell curing and the gene therapy.By replacing method,the AD rats showed signs of recovering to some extent on both histomorphology and behavior after transplanting NSCs into their brains.Transplanting NSCs along with the nerve nutrition factor (NTFs) showed better curative effects than NSCs transplantation alone.However,little is known about the molecular mechanism involving in the development of NSCs in vivo conditions.And the blindness of the treatment hindered the comparison of various affecting factors.The NSCs gene therapy is still in initial studying,with the effects of both cell replacement and gene therapy.This treatment genetically modified NSCs mainly by unitary nutrition fators such as nerve growth factor (NGF),brain-derived neurotrophic factor (BDNF),and glial cell line-derived neurotrophic factor (GDNF).And it was almost known nothing about the exogenous gene expression efficiency,the induce differentiation,the restore of function and security after genetically-modified NSCs transplanted into the AD rat brain.The detecting technology of NSCs transplanting curative effects of the AD rat is unitary at present.And the combined method is the developing trend,such as combining the immunohistochemical method with in vivo-tracking,and combining morphology index with the function index.

Objective To evaluate the clinical curative effect,survival rate and adverse reactions of three-dimensional conformal radiotherapy(3D-CRT )in combination with chemotherapy on lymph nodes metastasis of esophageal carcinoma.Methods Using supraclavicular 3D-CRT combined with chemotherapy on and simple 3D-CRT supraclavicular lymph node metastasis of esophageal cancer patients,3D-CRT com-bined synchronous chemotherapy (treatment group),51 cases,only 3D-CRT 49 cases (control group).3D-CRT combined synchronous chemotherapy 51 cases (treatment group),simple 3D-CRT 49 cases (control group).These patients 3D-CRT were given the total dose of 50 ~60Gy/25 ~30F.TN chemotherapy regi-mens were applied:paclitaxel 135 mg/m2 ,d1;Nedaplatin 25 mg/m2 ,d1,1 ~3,21 days cycle in fist week and fourth week.Results Local control and treatment group survival rates in 1,2 year were significantly higher than that of control group (P <0.05).Treatment group adverse reaction rate is higher than the con-trol group,but there was no statistically significant difference.Conclusions The recent curative effect and survival rate could be significantly improved by 3D-CRT joint TN synchronous chemotherapy regimen for pa-tients with supraclavicular lymph node metastasis of esophageal cancer,but the relatively high incidence of adverse reactions,clinical application should be considered comprehensively according to actual situation.

Objective To evaluate the effect of intrathecal methotrexate on activation of spinal astrocytes in a rat model of bone cancer pain (BCP).Methods Sixty female unmated Sprague-Dawley rats,aged 5-7 weeks,weighing 150-180 g,were randomly divided into 3 groups (n =20 each) using a random number table:sham operation group (group S),BCP group (group P),and BCP + methotrexate group (group PM).BCP was induced by injecting Walker-256 cancer cells into the medullary cavity of tibia.On day 7 after BCP,methotrexate 100 μg (diluted to 15 μl in artificial cerebrospinal fluid) was injected intrathecally over 10 s in group PM,and artificial ccrebrospinal fluid 15 μ1 was injected intrathecally over 10 s in S and P groups.The mechanical paw withdrawal threshold (MWT) was measured at 1 day before BCP and 3,7,14 and 21 days after BCP.Five rats were sacrificed after measurement of the pain threshold at 7,14 and 21 days after BCP,and the lumbar enlargement segments of the spinal cord were harvested for detection of the expression of glial fibrillary acidic protein (GFAP) by immuno-histochemistry.Five rats were sacrificed after measurement of the pain threshold at 14 days after BCP,and the expression of GFAP in the spinal cord was detected by Western blot.Results Compared with group S,the MWT was significantly decreased at 3,7,14 and 21 days after BCP in C and CM groups,the expression of GFAP was significantly up-regulated at each time point after BCP in group C,and the expression of GFAP was significantly up-regulated at 7 and 14 days after BCP in group CM (P＜0.05).Compared with group C,the MWT was significantly increased,and the expression of GFAP was significantly down-regulated at 14 and 21 days after BCP in group CM (P＜0.05).Conclusion The mechanism by which intrathecal methotrexate reduces BCP may be related to inhibition of spinal astrocyte activation in the rats.

Objective To investigate the risk factors for portal vein thrombosis (PVT)under anticoagulation therapy after surgery in patients with portal hypertension.Methods We made a retrospective analysis of clinical data of 96 portal hypertension patients for surgical treatment at our hospital.All the patients with postoperative PVT or without were divided into two groups.Risk factors that may predict PVT were analyzed.Results PVT developed in 41 (40.08%)of 96 patients after surgery.Risk factors such as sex,age,Child-Pugh classification,type of operation,portal pressure and the pressure difference before and after surgery,preoperative prothrombin time, preoperative platelet count,spleen index,and portal vein diameter were not predictors of PVT.However,splenic vein diameter was an independent risk factor for PVT (P = 0.036);postoperative PVT tended to develop when the splenic vein diameter was larger than 1 1 mm.Conclusion Preoperative color Doppler testing of splenic vein diameter can predict PVT after surgery in patients with portal hypertension.

Objective To provide a new approach of preservation for germplasm resources of Citrus medica var.sarcodactylis(CMS).Methods A micrografting test was made on CMS shoot-tips after vitrified cryopreservation,resulting in living shoot-tip.Results First the CMS shoot-tip is inoculated in medium consisting of MS,5%DMSO,and 5%sucrose for 48 h preincubation.Then the shoot-tips were cut off 3—4 mm long and fore-treated by 60%PVS_2 at room temperature(25℃) for 30 min.After that,they were treated by PVS_2 at 0℃for another 80 min and conserved in liquid nitrogen for 24 h.Next the shoot-tips were defrosted by aqueous bath at 40℃and cleaned twice at room temperature(25℃) by MT minimal medium with additive 1.2 mol/L sucrose,10 min once.Finally data collected recorded a higher survival rate of 81.25%by TTC inspectation and a better regeneration rate of 52.94%if the CMS shoot-tips had been grafted on darkly-cultured parental stock,growing,and differentiating normally afterwards.The plantlets detected by polymerase chain reaction(PCR) showed that the pathogens of Huanglong disease had been removed.Conclusion Therefore a conclusion is made that the approach of vitrified cryopreservation may be applied to the preservation of CMS germplasm resources.

Objective To lay a foundation for the cultivation of disease-free plantlet of Citrus medica L.var.sarcodactylis Swingle(CMS) by discussion of the factors affecting survival rate for micro-grafted shoot-tip of CMS.Methods With citrus limon seedlings cultured in the dark as the rootstock,shoot-tip of CMS was grafted to the inverse T-shaped cut in the cotyledon of rootstocks.Then an investigation was carried out on the factors which affected the survival rate for micrografted shoot-tip of CMS.The factors included age of rootstocks,scion size,culture medium with different concentrations of phytohormones,and rootstocks with or without cotyledon.Results The survival rate for micro-grafted shoot-tip of CMS was closely related to the factors mentioned above.A higher survival rate was obtained when CMS shoot-tip with 4 pieces of leaf primordia was grafted to the inverse "T" shaped cut in the cotyledon of parental stock(citrus limon seedlings) which was cultured in the dark for 14 days,and when the citrus limon seedlings were cultivated in MT-based medium with ?-naphthylacetic acid 0.1 mg?L-1 added.Conclusion The optimized micro-grafting conditions for CMS shoot-tips improve the propagation of micro-grafted plantlet and the survival rate of micro-grafted shoot-tip of CMS.

Objective To establish criteria for determination of sputum retention in elderly patients with non-ar-tificial airway. Methods Interview and Delphi methods were used to preliminarily determine clinical indication of sputum retentionand its classification through two-round expert consultation among 25 experts. Final criteria for de-termination of sputum retention were determined through experts group discussion. Results During the two rounds of consultation,all the questionnaires were collected,the experts' authority coefficients were 0.936 and 0.926 respec-tively,and the coordination coefficient of expert opinion was statistically significant.Criteria for determination of spu-tum retention included ineffective cough plus auscultation phlegm sound,with or without complaining of sputum,de-creased blood oxygen saturation,difficulty breathing and cyanosis. Conclusion Based on expert consultation and ar-gument,the criteria for determination of sputum retention can facilitate clinical nurses to detect sputum retention.

0 05).CONCLUSION:Na +-K +-ATPase is not sensitive to ischemic damage,but sensitive to reperfusion damage.Like hypothermia,scopolamine can protect Na +-K +-ATPase activity in the cerebral cortex during postischemic reperfusion,but hypothermia has no cooperating effect on scopolamine.