Abstract:ObjectiveTo construct a phage-displayed random combinatorial library of IgA affibodies and to analyze its directed in vitro evolution,so as to study the relationship of IgA affibody structure with its function.MethodsThe coding sequences of two affibodies,ZA1 and ZA2,were generated by overlapping PCR.The affibodies with a random linking peptide coding sequence in the 3′ terminal were randomly ligated and cloned into the KpnⅠsite of the phagemid pCANTAB5S to construct a combinatorial phage library.Totally four rounds of in vitro human IgA directed evolution were conducted,and selected phage clones were prepared individually to test the IgA binding activity by ELISA technology.ResultsThe combinatorial phage library was successfully constructed;it contained about 3.4×107 clones with a titer of 1.6×1012 TU/L,and the positive clones accounted for more than 79%.Sequence analysis showed that the two single affibodies were randomly linked by random linking peptides.The composition of the phage clones displaying three or four affibodies in tandem increased remarkably along with the rounds of selection,which indicated the successful IgA directed evolution.Three new arrangements of two,three or four affibodies in tandem were obtained.ELISA results demonstrated a significantly enhanced IgA binding activity of three or four affibodies in tandem.ConclusionA series of new IgA binding recombinants have been obtained by directed in vitro evolution of combinatorial phage library displaying randomly linked IgA affibodies.The three or four affibodies in tandem have much higher IgA binding activity than others,indicating that the in vitro molecular evolution is an effective way to produce IgA binding proteins with high activity.

Abstract:ObjectiveTo observe the effect of dexamethasone (Dex) and transforming growth factor β1 (TGFβ1) on the expression of plasminogen activator inhibitor-1 (PAI-1) in human ovarian cancer cell line HO-8910,and to explore its role in the development and progression of human ovarian cancer.Methods Human HO-8910 cells were treated with Dex(100 nmol/L) or TGFβ1(0.01-10 ng/ml) or both for different time periods,then the expression of PAI-1 mRNA and protein was examined by real-time PCR and Western blotting analysis.Act D was used to inhibit the synthesis of new mRNA so as to evaluate the stability of PAI-1 mRNA.ResultsTGFβ1 up-regulated PAI-1 mRNA expression in HO-8910 cells in a time- and dose-dependent manner,with the highest increase being 3.75 folds (P<0.01); Dex also up-regulated PAI-1 mRNA expression,with the highest increase being 1.65 folds (P<0.01).Dex combined with TGFβ1 markedly up-regulated PAI-1 mRNA level,with the highest increase being 12.7 folds (P<0.01),which was greatly higher than the sum of Dex and TGFβ1.Dex combined with TGFβ1 also greatly up-regulated the PAI-1 protein level compared with Dex or TGFβ1 alone.Dex,TGFβ1 or combination of them did not affect the stability of PAI-1 mRNA.ConclusionBoth Dex and TGFβ1 can up-regulate the expression of PAI-1 in HO-8910 cells,and the combination of them can work synergistically to up-regulate PAI-1 expression.The synergistic effect occurs at the transcriptional level,and it is not due to enhenced stability of PAI-1 mRNA.

Abstract:ObjectiveTo investigate relationship of GSTP1,RASSF1A polymorphisms and environmental agent with susceptibility to prostate cancer (Pca).MethodsThe GSTP1 and RASSF1A genotypes were determined by TaqMan/MGB Probe Technology in 103 patients with Pca and 103 normal controls. Multivariate logistic regression model was used to assess the association of smoking,alcohol drinking,tea drinking,weekly pork and beef consumption,and the genetic polymorphisms with the susceptibility to Pca,while taking into consideration of the environmental agent.ResultsThe frequencies of the GSTP1 AA,AG and GG genotypes were 66.02%,22.33%,and 11.65% in patients with Pca and 67.96%,29.13% and 2.91% in controls,respectively,with significant difference found between the two groups (χ2=6.35,P=0.04). The frequencies of RASSF1A CC,CA and AA genotypes were 88.34%,5.83%, and 5.83% in patients with Pca,and 85.44%,12.62%, and 1.94% in the controls,respectively,with no significant difference found between the two groups (χ2=4.63,P=0.10).Multivariate analysis showed a decreased risk in those who had a tea drinking history (OR=0.40,95%CI,0.19-0.82) and an increased risk in those who had a smoking history (OR=3.02,95%CI,1.44-6.32).ConclusionOur results indicate that GSTP1,RASSF1A polymorphisms are not associated with Pca susceptibility in Chinese Han nationality. Smoking is the risk factor of Pca,and tea drinking is a protective factor against Pca.

Abstract:ObjectiveTo investigate the relationship of abnormal insulin signal transduction with NF-κB activation in the adipose tissue of patients with type 2 diabetes mellitus(T2DM),and to study the effect of NF-κB decoy on insulin resistance in vitro.MethodsDecoy targeting NF-κB was designed and synthesized.Adipocytes from abdominal subcutaneous adipose tissues were obtained from T2DM patients and non-diabetic patients.The phosphorylation of insulin signal transduction molecules IRS-1 tyrosine and Akt-Ser473 of adipocytes were examined by immunoprecipitation and Western blotting analysis after stimulation with insulin.The activity of NF-κB in the adipocytes was analyzed by electrophoretic mobility shift assay (EMSA).The NF-κB decoy was transfected into the adipocytes of T2DM patients via liposome.ResultsThe phosphorylation levels of insulin signal transduction molecule IRS-1 tyrosine and Akt-Ser473 in adipocytes T2DM patients were significantly less than that in adipocytes from non-diabetes mellitus controls(P<0.05).The activity of NF-κB in adipocytes of T2DM patients was significantly higher than that in the controls(P<0.01).After transfection of NF-κB decoy,the activity of NF-κB was decreased compared with that before transfection(P<0.05),and the phosphorylation levels of IRS-1 tyrosine and Akt-Ser473 in adipocytes of T2DM patients were significantly increased compared with those before transfection(P<0.05).ConclusionInsulin resistance and excessive activation of NF-κB exist in the adipocytes of patients with T2DM; NF-κB decoy can partly ameliorate insulin resistance.

Abstract:1病例资料患者,男,24岁,2008年4月因尿毒症在我院行“亲属活体肾移植术”,手术顺利,术中留置F5输尿管支架管,术后患者常规使用免疫抑制剂治疗,各项指标正常。2个月后,膀胱镜检查发现患者膀胱内尿液浑浊,有大量白色絮状物,输尿管支架膀胱端钙盐沉着包裹。使用异物钳拔支架管,在约5 cm处遇较大阻力,患者感移植肾区疼痛不适,调换角度及力量后仍不能拔出。移植肾区超声检查提示：移植肾输尿管支架管周多发结石。抗感染治疗1周后,行体外冲击波碎石术（extracorporeal shock wave lithotripsy,ESWL）,术中分别对输尿管支架肾盂端、肾盂输尿管移行段及输尿管中段高密度区进行靶向重点碎石。术后再次行膀胱镜检查,术中感输尿管支架较前明显松动,可将输尿管支架牵引至尿道外口,但仍不能顺利拔出,遂换用适当的力量体外持续牵引输尿管支架管24 h。随后再次行ESWL,碎石过程配合外力牵引,变换调整碎石角度,逐渐将输尿管支架管完整拔出体外,见支架管周及管腔内附着大量结石。术后患者有轻度尿路刺激症状,抗感染补液治疗3 d后,患者症状完全消失,复查尿常规未见异常,顺利出院。

Abstract:ObjectiveTo investigate dihydroartemisinin(DHA)-induced apoptosis of human prostate cancer cells and the underlying molecular mechanism.MethodsNude mice were implanted with human prostate cancer PC-3 cells to establish tumor-bearing mouse model.Twenty models mice were evenly randomized into four groups:control group,solvent group(DMSO),large dose DHA (200 μmol/kg) group and low dose DHA (100 μmol/kg) group.The implanted tumors were observed on day 13 after drug administration.Morphological changes of PC-3 cells were observed by transmission electron microscope(TEM).The protein expression of apoptosis associated gene Bcl-2 and Bax were examined by immunohistochemical method.Apoptosis was detected by TUNEL assay.ResultsTEM examination revealed scattered apoptotic cells and apoptotic bodies in tumor tissues of mice in the DHA groups; immunohistochemical examination revealed that the protein expression of apoptosis-associated gene Bcl-2 was decreased and that of Bax was increased in DHA groups(P<0.05).TUNEL staining revealed that the rate of cell apoptosis increased significantly in DHA groups(P<0.05).ConclusionIt is demonstrated that DHA has strong effect in inducing apoptosis in human prostate cancer cell line PC-3 in vivo,which might be related to the down-regulation of Bcl-2 and up-regulation of Bax.

Abstract:ObjectiveTo study the effects of GSK-3β on the migration ability of hepatocellular carcinoma cells.MethodsInhibition tests with specific inhibitors of GSK-3 (LiCl and SB216763) and transfection with different plasmids(GSK-3β WT,GSK-3β S9A,and GID5-6) were used to alter the activity of GSK-3β.The migration ability of SMMC-7221 and Hep3B cells was observed using wound healing assay,transwell assay and the microtubule-organizing center (MTOC) test before and after treatment.Immunocytochemistry was used to investigate the distribution of phospho-GSK-3β.ResultsWound healing assay showed that LiCl and SB216763 decreased the migration of SMMC-7721 cells by 36.44% and 41.78%,respectively; and decreased the migration of Hep3B cells by both 26.66%.Transfection with GSK-3β WT increased the expression of GSK-3β and pGSK-3β,and the migration rate of SMMC-7721 cells decreased to 61.27%. Transfection with GSK-3β S9A increased GSK-3β expression and had no noticeable influence on pGSK-3β expression,and the migration rate of SMMC-7721 cells decreased to 38.61%.Transfection with GID5-6 had no noticeable influence on GSK-3β and increased pGSK-3β expression,and the migration rate decreased to 36.49%.Immunocytochemical staining showed that LiCl blocked pGSK-3β accumulation on cell edge as that in normally cultured cells.MTOC test showed that the MTOC positive rate was above 60% in Hep3B cells before adding inhibitors of GSK-3β,and it decreased after exposure to the inhibitors.ConclusionGSK-3β plays an important role in migration of hepatocellular carcinoma cells.Inhibitors of GSK-3β can affect the polarization and decrease the migration ability of hepatocellular carcinoma cells.

Abstract:Objective:To explore the role of sialic acid-binding immunoglobulin-like lectin-one (Siglec-1) in oxidized low-density lipoprotein (ox-LDL)-induced atherosclerotic inflammation.Methods:Ox-LDL was prepared by oxidization of native LDL;different concentrations of ox-LDL were used to treat U937 cells for 48 h.Cells and supernatants were collected.The expression of Siglec-1 protein and mRNA was examined by flow cytometry (FCM) and real-time quantitative RT-PCR,respectively.The levels of MIP-1α,MCP-1 and IL-8 in the supernatants were determined by ELISA.Results:Stimulation with ox-LDL significantly increased Siglec-1 protein and mRNA in U937 cells compared with the control group(P<0.01).Meanwhile,the levels of MIP-1α,MCP-1 and IL-8 in the supernatants were also significantly increased in a concentration-dependent manner compared with those in the control group(P<0.01).Conclusion:The proatherogenic effect of ox-LDL may be partly through up-regulating Siglec-1 expression in macrophages and enhancing the expression of inflammatory cytokines and chemokines.The activated macrophages recruit more macrophages and lymphocytes to the site of atherosclerotic plaques and exacerbate the inflammatory responses.

Abstract:ObjectiveTo establish a carotid arteriosclerosis model with New Zealand white rabbit by intravenous infusion of Helicobacter pylori(Hp),so as to lay a foundation for further investigating the relationship between Hp and carotid arteriosclerosis.MethodsEighteen New Zealand white rabbits were fed with high fat diet for six weeks;six of them were randomly chosen and sacrificed; the other twelve were evenly randomized into control group and experimental group.Animals in the experimental group were injected with 0.5 ml Sydney Strain 1(4×108 CFU) into the ear vein once a day for three days,and animals in the control group received normal saline in the same manner.All the animals were sacrificed on the eighth week.The blood lipid,carotid intima-media thickness (IMT),and plaque formation were observed before and 6,8 weeks after the intervention.The animals were sacrificed by air embolism and the carotid specimens were collected.The morphology of the blood vessels and the presence of plaque were observed with naked eye.H-E staining was used to observe the blood vessel diseases and intima thickness.ResultsAll the animals survived,and hyperlipidemia rabbit models were successfully established after 6-week feeding with high fat diet.The blood lipid level,carotid IMT,and blood vessel intima thickness were significantly increased in the experimental group compared with those in the control group at the eighth week（P<0.05）.More prominent atherosclerosis was noted in the experimental group compared with the control group on the eighth week.ConclusionCarotid arteriosclerosis model can be successfully established by intravenous injection of Hp in rabbits with hyperlipidemia.

Abstract:ObjectiveTo observe the expression of PPARγ in rat brain after status epilepticus. MethodsMale Sprague-Dawley rats were randomly divided into a control group and a status epilepticus group(n=10). Lithium-pilocarpine model was used to prepare epileptic model. Immunohistochemical method and Western blotting analysis were used to examine the changes of PPARγ distribution and expression in rat brain in the two groups.ResultsImmunohistochemical staining showed that the expression of PPARγ was weak in the brain of normal rats,and the expression was mainly on the neurons.The expression was strong in astrocytes after status epilepticus,and the expression was also noticed on cells with neuron shape.Western blotting analysis showed that the expression of PPARγ was greatly increased after status epilepticus.ConclusionThe expression of PPARγ is increased on astrocyte after status epilepticus in rats,which may have neuroprotective effect.This finding cast new lights on neuroprotective treatment for epilepsy.

Abstract:ObjectiveTo investigate the efficacy of 125 I in treatment of nude mice implanted with human glioma cell line U251 and the related mechanism.MethodsThe animal models of human malignant glioma were established with nude mice.125 I was implanted into tumors and the tumor growth was observed.The ultrastructural change was observed by electron microscopy.The apoptosis of the tumor tissue was examined by in situ end-labelling technique; expression of Bax/Bcl-2 was detected by flow cytometer.Blood vessel density and expression of VEGF,bFGF were observed by immunohistochemistry.ResultsNude mouse models of human malignant glioma were successfully established.Compared with control group,125 I significantly inhibited tumor growth in nude mice(P<0.05,P<0.01).Electron microscopy demonstrated that the tumor cells displayed apoptosis characteristics such as nuclear fragmentation,margination of condensed chromatin.The expression of Bax/Bcl-2 in the 125I implanting group (1.88±0.47)was significantly increased compared with that in the control group (1.11±0.52,P<0.01),and the apoptosis rate of tumor tissues in the 125I implanting group (3.64±0.89) was significantly higher(0.81±0.45,P<0.01).The blood vessel density in the 125I implanting group was significantly lower than that in the control group (1.40±0.55 vs 3.23±0.87,P<0.01).The expression of VEGF,bFGF in 125I implanting group was significantly lower than that in the control group (1.58±0.55 vs 4.19±0.45 for VEGF,P＜0.01; 2.44±0.89 vs 3.52±0.79 for bFGF,P<0.05).Conclusion125I has prominent inhibitory effect against human gliomas,and the main mechanism might be the promotion of apoptosis and decrease of blood vessel density.

Abstract:ObjectiveTo observe the influence of hepatocyte growth-promoting factor (pHGF) on monocyte chemotatic protein-1-(MCP-1) and aristolochic acid Ⅰ(AAⅠ)-induced epithelial-to-mesenchymal transition (EMT) and apoptosis of human kidney epithelial cell line(HKC).MethodsThe HKC cells were randomly divided into blank control group (control groups), epithelial-to-mesenchymal transition model group (model group),and pHGF inhibition group (pHGF groups) with pHGF at different concentrations (0.15,1.5,15,150,and 1 500 ng/ml).The EMT model was established by exposing HKC cells to MCP-1(0.1 μg/ml)and AAⅠ(10 μg/ml).Cells in the pHGF groups were the model cells treated with different concentrations of pHGF.Cells in the control group were cultured routinely.WST-8 method and flow cytometry were used to observe the proliferation and apoptosis of HKC cells,respectively.The mRNA expression of α-smooth muscle actin(α-SMA) was determined by reverse transcriptase-polymerase chain reaction(RT-PCR),and the expression of α-SMA,fibronectin (FN),and transforming growth factor-β1(TGF-β1) in HKC cells were assessed by indirect enzyme immunohistochemistry.ResultsThe cell inhibitory rate,apoptotic rate,and expression of α-SMA mRNA were significantly increased in the model group and pHGF groups compared with those in the control group (P<0.01),indicating the successful establishment of EMT model.Compared with the model group,pHGF at 150 ng/ml,but not at other concentrations,significantly decreased the inhibition rate of HKC cells(P<0.01).The apoptotic rate of HKC cells in all the pHGF groups were significantly lower than that in the model group (P<0.01).pHGF at 150 ng/ml also greatly decreased the expression of α-SMA mRNA,and significantly down-regulated the expression of α-SMA,TGF-β1,and FN protein.ConclusionpHGF at 150 ng/ml can partly reverse MCP-1- and AA Ⅰ-induced HKC cell growth inhibition,apoptosis,and EMT.

Abstract:ObjectiveTo study the effect of ischemic preconditioning on heat-shock protein 70 (HSP70) expression and the learning,memory functions after forebrain ischemia-reperfusion injury in gerbils.MethodsGerbils (n=100) were evenly randomized into four groups:Sham group,ischemia-reperfusion(I/R) group,ischemia preconditioning(IP) group,and Cycloheximide + IP group (Cycloheximide was administered 30 min before IP).Transient forebrain ischemia-reperfusion model was established by bilateral common carotid artery occlusion according to the method described previously,and ischemia preconditioning model was established as described by Kitagawa.Changes of neuron morphous in hippocampus CA1 region were observed by H-E staining 1,2,and 3 days after reperfusion.The expression of HSP70 was examined by immunohistochemistry and Western blotting assay.Neuron apoptosis was detected by TUNEL method.The learning/memory functions of gerbils were examined using 4-PTT dry path maze 3,4,5,6,and 7 days after reperfusion.ResultsCompared with Sham group,I/R group had significantly decreased survival neurons(P<0.05),increased neuron apoptosis(P<0.05),increased expression of HSP70 (P<0.05),and decreased learning/memory functions(P<0.05).Compared with I/R group,IP group had significantly increased survival neurons(P<0.05),decreased neuron apoptosis (P<0.05),and increased expression of HSP70 (P<0.05),and improved learning/memory functions(P<0.05).However,cycloheximide almost totally abolished the effect of ischemia preconditioning,with the neuron morphology,density,apoptosis,HSP70 expression,and learning/memory functions similar to those of I/R group.ConclusionIschemic preconditioning can protect against cerebral ischemia injury and improve the learning/memory functions after forebrain ischemia-reperfusion damage,which is possibly through promoting HSP70 expression and starting endogenous neuroprotective mechanism,subsequently reinforcing the protective effect against ischemia.

Abstract:ObjectiveTo study the clinicopathologic properties of hippocampus glioma with temporal lobe epilepsy(TLE) as the main manifestation.MethodsThe clinical data of 6 patients with TLE and hippocampus masses were retrospectively analyzed.The 6 patients included 2 males and 4 females, with the longest history of epilepsy being 4 years.ResultsAll the 6 patients were surgically treated and were pathologically confirmed to have gliomas,including one angiocentric glioma(AG),one pilocytic astrocytoma(PA) with mucous degeneration,one astrocytoma,one oligoastrocytomao,one oligodendroglioma and one glioblastoma.Glial fibrillary acidic protein(GFAP) was strongly positive in all cases.Epilepsy disappeared after surgery in 5 cases; one patient had recurrence and received a second operation.ConclusionHippocampus gliomas with TLE as the main manifestation are usually low-grade gliomas of various types,and operation can obtain satisfactory prognosis.

Abstract:ObjectiveTo summarize our experience on laparoscopic radical prostatectomy via single incision in treatment of early localized prostate cancer.MethodsFrom June 2009 to August 2009,five patients with localized prostate cancer（T1c）received laparoscopic radical prostatectomy via single incision.A home-made multichannel port was inserted extraperitoneally through a 3 cm incision under the umbilicus.A 10 mm TROCAR and two 5 mm TROCAR were inserted.The prostate was isolated and excised,then the bladder urethral anastomosis was performed.ResultsThe procedures were successful in all the five cases with no transversion to open or standard laparoscopic approach.The mean operating time,the mean operative time for prostate excision,and the mean time for urethrovesical anastomosis were (167±31.5) min(ranging 135-210 min),(115±26) min (ranging 90-150 min),and (52±5.7) min(ranging 45-60 min),respectively.The estimated blood loss averaged (90±62) ml (ranging 50-200 ml).Positive margin occurred in one case.Transient incontinence occurred in two cases after the catheter was removed one week later.All patients had a prostate-special antigen level ＜0.2 μg/L during a follow-up of 4-12 weeks.ConclusionLaparoscopic radical prostatectomy via single incision is feasible and safety.Excellent instruments and skilled surgeon are the keys for the success of the operation.

Abstract:ObjectiveTo evaluate the shape and texture of the alveolar bones before dental implantation using three-dimension reconstruction techniques by multi-slice spiral CT (MSCT).MethodsSeventeen patients, whose alveolar condition was not clear enough to receive dental implantation, were screened out using conventional dental periapical and panoramic X-ray examination.The occlusion cast was made and a radiological guide template with titanium tube was prepared in the implantation area.The mandibular and maxillary bone was scanned with MSCT.The height,width and density of the alveolar were measured through three-dimensional reconstruction,so as to determine whether the alveolar was suitable for dental implant surgery.Implantation using ITI dental implant system was performed in patients whose alveolar condition met the implantation criteria.ResultsPre-operation MSCT examination indicated that two cases were not suitable for the surgery due to a width of alveolar bone less than 5.3 mm and a height of the residual bone under the maxillary sinus less than 3 mm.Fifteen patients suitable for the surgery underwent successful implantation.ConclusionThe three-dimensional reconstruction of MSCT images of the jaws can provide more comprehensive and reliable information of the non-security areas and mandibular or maxillofacial alveolar bone defects before implantation.

Abstract:ObjectiveTo observe the short-term clinical efficacy and adverse effects of paclitaxel plus pirarubicin(THP) or epirubicin(EPI) in the neoadjuvant chemotherapy of locally advanced breast cancer (LABC).MethodsForty patients with newly diagnosed LABC were enrolled and were randomly assigned to two neoadjuvant chemotherapy groups(n=20):PT group received THP plus paclitaxel,and PE group received EPI plus paclitaxel.The tumor size,the pathological response,and the adverse reactions were observed on a weekly basis.ResultsAll the patients received six courses of chemotherapy.Thirty-nine of the patients received 2-4 cycles of neoadjuvant chemotherapy before they received operation; the remaining course of treatment was carried out after surgery.One patient received operation 6 weeks after neoadjuvant chemotherapy.The pathological complete response (pCR),clinical complete response (cCR) and partial response (cPR) in PT group were 10.0%,20.0% and 75.0%,respectively,and in PE group were 5.0%,30.0% and 70.0%,respectively.There was no significant difference in the total response rates (RR) between the two groups.Cardiac toxicity and leukopenia in the two groups were grade Ⅰ-Ⅱduring the chemotherapy. The incidence of grade Ⅲ gastrointestinal reaction (nausea and vomiting) was 45.0% in the PT group and 90.0% in the PE group (P<0.05).ConclusionNeoadjuvant chemotherapy with paclitaxel plus THP or EPI has satisfactory short-term clinical efficacy and adverse effects for LABC.The rate of radical surgical resection after chemotherapy is high.The gastrointestinal adverse reaction in the PT group is significantly lower than that in PE group.Cardiac toxicity and leukopenia are weak and bone marrow suppression is slight; PT regimen is improved compared with PE group.

Abstract:ObjectiveTo propose an effective algorithm for solving the shortest circuit problem of military health support.MethodsPartheno-genetic algorithm (PGA),which only used mutation operation and selection operation,was adopted in the present study.The algorithm was based on K-random-nearer-neighbor algorithm combined with two-random-point exchange,two-neighbor-point exchange,circular-based part inversion and random insertion mutation operations.Furthermore,greedy strategy was applied in selection to improve the hill-climbing capability of PGA.ResultsThe simulation results of CTSP31 and standard dataset from TSP library indicated that the PGA was more effective than existing algorithms from the literature.ConclusionPGA can serve as a basis for further development of a computer-assisted program,and it provides optimized decision-making scheme for improving the quality and speed of military medical service disposition.

Abstract:Objective:To prepare a simple,practical fiberoptic bronchoscope (FB) training simulator,and to evaluate its training efficacy.Methods: A box apparatus was designed for basic and advanced FB training; the box consisted of five boards with holes in it.The holes were aligned according to the entries of bronchus to pass the FB.Assistant materials could be placed in the box to simulate foreign bodies due to diseases,which was aimed to help the training of sophisticated skills including tracheal tube intubation,biopsy,brushing and bronchoalveolar lavage.To testify the training efficacy,forty volunteer residents who had never manipulated a FB were randomly assigned into two groups:one group was trained using the prepared training box (Group TB,n=20) and the other group was trained using video-based technique (Group C,n=20).The trainees of the two groups received an intubation examination using an advanced patient simulator, and the periods of intubation was compared between the two groups.Results: Flexible fiberoptic bronchoscope could smoothly pass through the holes in the box; the photos of the holes under bronchoscopy well simulated the normal images.All residents completed the training and examination.The mean duration of intubation was (55.7±10.2) s in Group TB and (69.3±8.9) s in Group C (P<0.001).Conclusion: The present training box is simple and practical,and it can be used for advanced training of FB,with obvious training efficacy.

Abstract:ObjectiveTo study the influence of recombinant human erythropoietin (rhEPO) preconditioning on the liver function and expression of nuclear factor-κB (NF-κB) during early stage following liver transplantation,and to investigate the possible mechanism of rhEPO preconditioning on ischemia-reperfusion injury after liver transplantation.MethodsTwenty-six patients with advanced hepatic cirrhosis were randomly divided into two groups(n=13): the rhEPo pre-treatment group received subcutaneous injection of rhEPO 100 U/kg at 1,3 and 5 d before liver transplantation,and the control group received 2 ml normal saline in the same manner. The peripheral blood samples were harvested at 1,2,4 and 6 h after blood supply recovery in the donator liver to examine the hepatic functions. The NF-κB p65 expression in the peripheral blood samples were examined by Western blotting analysis,the TNF-α level in the blood was detected by ABC enzyme linked immunosorbent assay. Serum ALT and AST were also determined.ResultsThe liver function indices and the levels of serum NF-κB p65,TNF-α in the rhEPO pre-treatment group were significantly lower than those in the control group (P<0.05).ConclusionPre-treatment with rhEPO can inhibit hepatic inflammation early after liver transplantation,protecting hepatic function and reducing ischemia-reperfusion injury after liver transplantation.

Abstract:ObjectiveTo observe the expression of Toll-interacting protein (Tollip) in appendix and analyze its significance during acute inflammation.MethodsThirty-three patients with acute appendicitis were included in the present study and 6 subjects with non-inflammatory appendixes were taken as controls (without inflammatory changes).The pulse rate,body temperature (BT),white blood cell (WBC) count and neutrophil (NEUT) count were observed one hour before appendectomy.Based on H-E staining and pathological examination,the appendix samples were divided into four groups:non-inflammatory appendix (A),simple appendicitis (B),suppurative appendicitis (C) and gangrenous appendicitis (D).The expression of Tollip protein (the localization,qualitative and semiquantitative analysis) was analyzed using immunohistochemistry and digital image analysis.The correlation of Tollip with pusle rate,BT,WBC and NEUT was also analyzed.ResultsSignificant differences in BT,WBC and NEUT were found between different pathological groups (P<0.05).Tollip protein was mainly expressed in the epithelium mucosa and glandular epithelium of appendix,but was not detected in the neutrophils.During the development of appendiceal inflammation (non-inflammatory appendix to simple appendicitis,to suppurative appendicitis,then to gangrenous appendicitis),the expression of Tollip protein gradually increased and was significantly correlated with WBC (P<0.05).ConclusionDuring the development and progression of acute appendicitis,Tollip expression is up-regulated in appendix tissues and is closely correlated with systemic responses of inflammation,such as the increase of WBC.

Abstract:ObjectiveTo establish an HPLC method for simultaneous determination of syringin,schizandrin,deoxyschizandrin and schisandrin B in Shuangwu capsules. MethodsThe HPLC method was employed using a Diamonsil C18 column (200 mm×4.6 mm,5 μm) with a mobile phase of methanol-acetonitrile (1∶1,A) and water (B). The gradient elution program was as follows: 0-5 min,35%-60%A; 5-10 min,60%-70%A; 10-50 min,70%-90%A; 50-90 min,90%A. The flow rate was 1 ml/min. The detection wavelength was set at 220 nm and the temperature was at 35℃.ResultsThe linearity was obtained within the range of 1.28-20.40 μg/ml for syringin (r=0.999 7),6.30-100.80 μg/ml for schisantherin (r=0.999 6),1.20-19.20 μg/ml for deoxyschizandrin (r=0.999 8),and 3.75-60.00 μg/ml for schisandrin B (r=0.999 6). The RSD values of precision were less than 1% for all the four components. The results showed that the samples were stable in the room temperature for at least 24 h. The average recovery rates of syringin,schizandrin,deoxyschizandrin and schisandrin B were 99.47%,102.50%,99.21% and 101.86%,respectively. Conclusion Our method is rapid,easy to perform and accurate; it can be used to control the quality of Shuangwu capsules.

Abstract:Controlling serum phosphorus levels is critical in patients with renal failure.Currently phosphate-binding agents are widely used to reduce phosphate absorption in patients with end-stage renal disease.If possible,serum phosphorus level should be reduced without disturbing calcium homeostasis or increasing accumulation of potentially toxic elements.Aluminum hydroxide and traditional calcium-based phosphate binders are commonly used to control serum phosphorus level.Aluminum hydroxide can effectively lower serum phosphorus level,but aluminum can accumulates in the body and results in toxic effect.Traditional calcium-based phosphate binders tend to promote hypercalcemia and calcium overloading,and accelerate cardiovascular calcification.Therefore aluminum-free and calcium-free phosphate-binding agents have become the focus of study; however,agents like sevelamer hydrochloride and lanthanum carbonate are not widely used due to high price,although they are effective in controlling serum phosphorus level.New generation of phosphate binders,such as colestilan,nicotinic acid and magnesium salt, are cheaper than their previous counterparts,but their long-term effect still needs to be observed.This article summarizes the progress of non-calcium phosphate binders in treatment of end-stage renal diseases,hoping to help clinical drug usage.

Abstract:Id proteins,or inhibitors of differentiation/DNA binding,are negative regulators of helix-loop-helix(HLH) type transcription factor,and they can inhibit cell differentiation and promote cell proliferation.Many experiments have proven that high expression of Id proteins in tumors was significantly related to malignant behaviors of tumors,such as tumor invasion and metastasis,suggesting Id proteins can be used to predict the prognosis of tumors.Abnormal expression of Id protein in malignant tumor might be associated with the invasive phenotype and poor prognosis of tumors.Id protein-targeted therapy can effectively inhibit tumor proliferation,invasion and metastasis.This paper reviews the mechanism by which Id protein influences the prognostic factors,such as malignant proliferation,invasion and metastasis,angiogenesis and the researches on progress of related clinical researches.

Abstract:ObjectiveTo design and prepare intensive training models for key steps of complicated laparoscopic urology surgery,and to use them for training of new learners.MethodsBased on the key steps for laparoscopic surgery in urology,such as radical prostatectomy,nephron-sparing surgery (NSS) and pyeloplasty for ureteropelvic junction obstruction (UPJO),we prepared the models of retropubic vesicourethral anastomosis,partial nephrectomy and pyeloplasty UPJO using intestines of pigs and chickens,kidney of pigs,swim bladder and ureter of pigs,etc.The models were used to train the new learners and the training outcomes were observed.ResultsThe models of retropubic vesicourethral anastomosis,partial nephrectomy and pyeloplasty UPJO were successfully prepared,and they satisfactorily simulated various procedures; they costed less,were easy to make,and had rich resources.The models were suitable for training of new learners.After intensive training with the models,the new learners could master the skills and became more self-confident.ConclusionWe have successfully prepared the models for intensive training of the key steps of complicated laparoscopic surgery in urology; the model can meet the demand for training of the new learners.

Abstract:Liver transplantation is the only definitive treatment modality of end stage liver diseases.Demyelinative disease is rarely seen in patients after liver transplantation,but a higher incidence has been noticed recently.The disease is liable to be misdiagnosed as immunosuppressant-induced psychiatric disorders at early stage.Central pontine myelinolysis is more disastrous and it will greatly influence the short-term survival and long-term life quality of the patients after liver transplantation.When it is manifested as peripheral nervous system disorder,the disease usually has a subtle onset,making it difficult for diagnosis and treatment.The causes for various demyelinative diseases should be understood to prevent it in patients after liver transplantation.The specific mechanism for demyelinative disease after liver transplantation remains unclear.We introduce the possible causes of common demyelinative diseases,hoping to provide reference for prevention and treatment of such conditions after liver transplantation.

Abstract:目的观察TGFβ1基因codon10(Leu＞Pro)的单核苷酸多态性(SNP)对肝脏细胞功能的影响。方法构建含codon10(Leu＞Pro) 的TGFβ1(LAP＋成熟态单体)真核表达重组体CMV-Leu、CMV-Pro。在HepG2、LX-2细胞中转染pcDNA3.1空载体、CMV- Leu、CMV-Pro,培养24 h后,ELISA检测细胞培养上清液中TGFβ1的分泌量。MTT实验观察HepG2细胞转染后的增殖情况,流式细胞术检测LX-2细胞转染后CD83的表达。结果转染CMV-Leu和CMV-Pro能增加HepG2、LX-2细胞的TGFβ1分泌量,且转染CMV-Pro的细胞TGFβ1分泌量比转染 CMV-Leu的细胞高(P＜0.05)。转染CMV-Pro能明显增高HepG2细胞的增殖活性(P<0.01),而转染CMV-Leu能明显增加LX-2细胞CD83的表达率(P<0.01)。结论TGFβ1基因codon10(Leu＞Pro)的SNP对肝脏细胞TGFβ1的分泌、细胞增殖活性及CD83的表达具有明显影响。

Abstract:目的研究母鼠怀孕中后期地塞米松(DEX)暴露对子代大鼠发育过程中痛觉敏感性的影响。方法以母鼠孕期中后半程(9～12 d)注射DEX(每天100 μg/kg)和注射生理盐水后出生的大鼠为样本,通过足底部痛阈测定,观察子代大鼠出生后3 d及1、2、3、4、5、6、7、8周时机械痛阈和热痛阈的变化。结果两组子代大鼠的外周机械痛阈和热痛阈均随着年龄的增长逐渐增加。出生后3 d和1周时,DEX组机械痛阈显著低于对照组(P<0.05),而2周和3周时,DEX组机械痛阈明显著高于对照组(P<0.05),4周以后两组间无显著差别。对于热痛阈,大鼠出生后3 d DEX组显著低于对照组(P<0.05),而出生后1周及以后的时期两组间无显著差异。结论母体孕中后期注射DEX可以增强子代大鼠发育早期的痛觉敏感性,而这种影响可能是短暂性的。

Abstract:1病例资料患者,男,73岁,因反复活动后胸闷气促10余年,突发胸闷、大汗4 h于2009年1月19日急诊收治入院。既往高血压病史10年,长期服用福辛普利 (蒙诺)、美托洛尔(倍他乐克)等药物,血压控制尚可。入院查体：BP 130/75 mmHg（1 mmHg=0.133 kPa）,神志清,双肺呼吸音清晰,未闻及明显干湿性啰音。心率84次/min,心界向左下扩大,各瓣膜听诊区未闻及病理性杂音。腹软,肝脾未及。双下肢无水肿。心电图示：窦性心律,完全性左束支传导阻滞,V1～V5导联ST段呈缺血性改变。肌钙蛋白I：0.38 μg/L。2009年1月22日在局部麻醉下行冠状动脉造影术,结果显示：左主干远端30%狭窄,前降支近段狭窄60%,分出第一对角支后狭窄70%,回旋支未见明显狭窄,右冠中段次全闭塞。遂对该病变处行经皮腔内冠状动脉成形术(PTCA)+支架植入术,植入直径4.0 mm、长度36 mm的乐普支架,手术顺利。术后第2天17：50患者突发胸闷不适,伴冷汗,无创血压监护BP 84/56 mmHg,查体：双肺呼吸音粗,双肺底可闻及细湿啰音；心率100次/min,律齐,心界向左下扩大,各瓣膜听诊区未闻及病理性杂音,无心包摩擦音。心电图示较前无明显变化,结合病史考虑急性左心衰竭引起的低血压,予吗啡、呋塞米（速尿）纠正心衰,多巴胺5 μg/（kg·min）维持血压治疗；22：30 BP 72/55 mmHg,心率110次/min,多巴胺增量至10 μg/（kg·min）,并加用间羟胺1.33 μg/（kg·min）。第3天患者仍反复感胸闷不适,无创血压监护交替测量两侧肱动脉压波动于70～90/40～50 mmHg,多巴胺逐渐增量至30 μg/（kg·min）,间羟胺增量至4 μg/（kg·min）,血压仍维持不住,逐渐降低,加用肾上腺素0.17 μg/（kg·min）。此时患者出现全身湿冷,桡动脉搏动细弱,脉氧测不出,心率160~180次/min。11:20急查血气分析：pH 7.289,PCO233.1 mmHg,PO2135.8 mmHg,HCO3- 17.3 mmol/L；16:05复查血气分析：pH 7.266,PCO231.2 mmHg, PO2 113.9 mmHg,HCO3- 16 mmol/L。在抽取股动脉血行血气分析时发现股动脉搏动较强,为精确监测患者动脉血压,于16:30行右侧股动脉穿刺,置入6F动脉鞘管,进行有创动脉压监测,显示股动脉压力134/58 mmHg,心率140次/min,即停用肾上腺素,多巴胺减量至20 μg/（kg·min）,间羟胺减量至2.67 μg/（kg·min）；23：00 BP 128/63 mmHg,心率105次/min,停用间羟胺,多巴胺逐渐减量至2 μg/（kg·min）。3 d后拔除动脉鞘管,停用有创血压监测。2009年2月22日患者顺利出院。