Abstract:Objective:To search for a method to induce human embryonic stem cells (hESCs) differentiating into neural stem cells (NSCs) in vitro. Methods:HESCs were induced to differentiate into NSCs by three-step differentiation under a condition simulating the microenvironment and different development stages of neural cells in vivo. The surface markers of hESCs and NSCs were detected by morphological observation, immunocytochemistry assay, flow cytometry and RT-PCR. The plasticity of NSCs was evaluated by differentiating test.Results:The hESCs retained expression of SSEA-4, TRA-1-81 proteins and Nanog genes after cultured for 50 passages. Flow cytometry revealed the positive rate of SSEA-4 was 83.44%.After induced by the three-step differentiation the purity of nestin-positive cells was higher than 90%. Flow cytometry revealed that the positive rate of nestin was 89.38%. The differentiated cells retained the characteristics of NSCs after repeated passaging and could be further induced into neurons, astrocytes and oligodendrocytes. Conclusion:The present three-step differentiation method can induce hESCs into high purity NSCs, while retaining the plasticity of stem cells.

Abstract:Objective:To study the effect of genetic polymorphisms of cytochrome P4501A1 (CYP1A1), N-acetyltransferase 2 (NAT2) on the susceptibility to renal cancer.Methods: The genetic polymorphisms of CYP1A1 and NAT2 were examined in the renal cancer patients and controls using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) methods.Results: The distribution of CYP1A1 (W/M) and NAT2 (intermediate) genotypes was significantly different between the renal cancer patients and controls (P<0.05).Individuals carrying CYP1A1 (W/M) or NAT2 (intermediate) genotypes had an increased risk for renal cancer（OR=2.487, 95% CI: 1.493-4.142; OR=1.970, 95% CI:1.128-3.442，respectively). Multivariate analysis showed increased risk for renal cancer patients carrying CYP1A1 (W/M) or NAT2 (intermediate) genotypes and those who smoke.Conclusion: The genotypes CYP1A1 (W/M) and NAT2 (intermediate) are the risk factors of renal cancer, and the 2 genotypes have a interactive effect and both have a joint effect with smoking.

Abstract:Objective:To investigate the inhibitory effects of RNA silencing via adenovirus-mediated vascular endothelial growth factor receptor (VEGFR) shRNA on proliferation of lung adenocarcinoma cells in vitro and in vivo．Methods: Ad-VEGFRshRNA adenovirus containing enhanced green fluorescent protein (EGFP) gene and VEGFRshRNA was constructed and was used to infect A549 cells; fluorescent microscopy was used to observe the infection efficiency. Western blotting assay was used to examine the expression of VEGFR protein in A549 cells. MTT method was used to examine the cell viability and the cell growth curve was drawn. The inhibition of cell growth was examined by cell cycle and colony-forming test. Meanwhile, nude mice were transplanted with A549 cells to establish tumor-bearing model, and the long term growth of tumor was observed.Results: Western blotting revealed that the expression of VEGFR was obviously decreased in the RNA interference group. The cell growth curve indicated that the cell growth was obviously inhibited after RNA interference. Cell cycle and colony-forming test indicated that the tumor growth was obviously inhibited after RNA interference. In vivo study with nude mice also indicated that RNA interference obviously inhibited tumor growth.Conclusion: The constructed VEGFR-targeted shRNA can effectively inhibit VEGFR expression in A549 cells and can suppress the growth of A549 cells.

Abstract:Objective:To establish a lung cancer phage-peptide library and to screen for biomarkers for early detection of lung cancer.Methods: A phage display library was constructed using 30 lung-cancer tissue samples from Changhai Hospital. Protein-A/G agarose was used to enrich IgG from control sera as well as lung cancer sera. Five biopannings were carried out for enrichment of lung cancer-specific phage clones. Five hundred phage clones were randomly selected,those with D>2.0( lung cancer plasma pool v.s. control plasma pool)were selected for DNA sequencing and protein prediction. Results: (1) The recombination rate of the phage library was 60%,with the average phage titer being 3.0×106 pfu and a volume being 9.0×106 pfu.(2) Of 19 phage colonies selected by ELISA and sequenced,9 were cancer-related genes,8 with unknown function,and 2 were not related to cancer. Conclusion: The screened genes in the phage colonies might serve as biomarkers for the early diagnosis of lung cancer.

Abstract:Objective:To construct a GPC3 green fluorescent protein eukaryotic expression vector pEGFP-N2-G-PC3,and analyze its effects on the growth promoting effect of growth factors (fibroblast growth factor-2，FGF2; insulin-like growth factor-2 ，IGF2; transforming growth factor-β1，TGF-β1; and bone morphogenetic protein-4，BMP4) in human hepatoma cell line GPC3-SK-Hep-1.Methods: A eukaryotic expression vector for GPC3 genes (pEGFP-N2-GPC3) was constructed by recombinant DNA technique and was transfected into SK-Hep-1 cells by LipofectamineTM2000; the cells stably expressing GPC3 were screened out by G418 (600 μg/ml).The mRNA expression of GPC3 was detected by RT-PCR method and the protein expression of GPC3 by Western blotting and fluorescence microscope.Effects of GPC3 gene on the growth promoting effects of the above growth factors were examined by MTT.Results: The recombinant plasmid was verified to be correctly constructed by restriction endonuclease analysis and DNA sequencing.The green fluorescence was detected in the transfected SK-Hep-1 cells under fluorescence microscope.RT-PCR and Western blotting both confirmed that GPC3 was successfully expressed in SK-Hep-1 cells.FGF2-induced cell proliferation was significantly decreased by GPC3 gene,whereas the growth promoting effects of IGF2,TGF-β1 and BMP4 were not altered by GPC3 gene.Conclusion: We have successfully obtained the synthetic GPC3 protein,

Abstract:Objective:To investigate the effects of 5α - dihydrostestosterone (DHT) on calcium mobilization and growth of prostate cancer cell line LNCaP.Methods: Intracellular calcium concentration (\[Ca2+\]i) was assayed by MiraCal Image System using Fura-2/AM as Ca2+ fluorescence probe. Cell viability was observed by MTT assay and apoptosis by flow cytometry. Results: The calcium levels rapidly increased following addition of DHT, with the latency of response only in seconds. DHT at the concentrations of 1, 10, 100 and 1 000 nmol/L increased \[Ca2+\]i from (28±5), (29±5), (28±4) and (28±9) nmol/L to (31±3) ( P>0.05,65±9) (P<0.01), (193±33) (P<0.001) and (208±42) nmol/L (P<0.001), respectively. The response induced by 1 000 nmol/L DHT was similar to that induced by 100 nmol/L DTH. DHT 1 000 nmol/L did not increase \[Ca2+\]i under extracellular Ca2+ -free condition. Blockers of L-type voltage-gated calcium channels, including verapamil (50 μmol/L), diltiazem (100 μmol/L) or nifedipine (5 mmol/L) at 37℃ for 5 min prior to stimulation with 1 000 nmol/L DHT, completely inhibited DHT-induced \[Ca2+\]i rise. Pre-treatment with inhibitor of phospholipase C such as neomycin sulfate (1 mmol/L) at 37℃ for 3 min or inhibitor of ryanodine receptor such as procaine (50 mmol/L) at 37℃ for 3 min had no influence on \[Ca2+\]i rise induced by 1 000 nmol/L DHT. The optical density (D) values and early apoptosis rates of the cells stimulated with 1 000 nmol/L DHT for 48 h were significantly different from those of cells pre-treated with verapamil prior to stimulation with 1 000 nmol/L DHT (\[0.67±0.10\] % vs \[2.13±0.16\] % and \[14.31±2.29\] % vs \[1.07±0.19\] %,P<0.01).

Abstract:Objective:To observe the influence of minocycline on expression of NF-κB, GFAP, and IL-1β in rats with vascular dementia，so as to study the neuroprotective mechanism of minocycline for vascular dementia.Methods: An animal model of vascular dementia was established by chronic bilateral common carotid artery occlusion (BCCAO). Animals were randomly divided into sham-operation group(S), 4-week model group (M4), 8-week model group (M8), 16-week model group (M16), 4-week model + Minocycline group (T4), 8-week model+Minocycline group (T8), and 16-week model+Minocycline group (T16). The behaviors of animals were tested with Morris water maze and shuttle box task. Expression of NF-κB and GFAP was measured by enzyme linked immunosorbent assay (ELISA), immunohistochemistry and Western blotting,and IL-1β by ELISA.Results: Minocycline greatly improved the behaviors of mice with vascular dementia, and promoted the learning, memory and responding abilities. The expression of NF-κB, GFAP and IL-1β in all the model groups and Minocycline treatment groups were significantly higher than that in the sham-operation group (P<0.01); and those of Minocycline treatment groups were significantly lower than the corresponding model groups (P<0.01).Conclusion: Minocycline can decrease the expression of NF-κB, GFAP, and IL-1β in the brain of rats with vascular dementia, and protect brain by inhibiting the activation of astrocytes and neuroinflammation.

Abstract:Objective:To study the expression of neuroglobin (Ngb) in rat cerebral ischemia model and the neuroprotective effect of Ngb after ischemia and hypoxia. Methods: Totally 113 Sprague-Dawley rats were randomly divided into sham-operated group, middle cerebral artery occlusion (MCAO) group, hemorrhagic infarction (HI) group and hemin treatment group. The brain water content, infarcted tissue volume, neuropathologic changes (H-E staining) and expression of Ngb (immunocytochemical staining) were examined 3, 6, 12, and 24 h after model establishment. Results: The brain water contents and the infarcted tissue volumes in the hemin treatment group were significantly different from those of the MCAO group and HI group (P<0.01). The brain edema was obviously increased in HI group at 12 h. Neuropathologic examination showed that there were fewer necrotic neurons, milder edema and stronger Ngb expression in the Hemin treatment group than in the MCAO group and HI group. Immunocytochemical staining showed that the Ngb positive neurons in Hemin treatment group were more than those in the MCAO and HI groups. Conclusion: Earlier peak of brain edema may lead to aggravation of disease. Hemin-induced Ngb expression may relieve brain damage during focal cerebral ischemia.

Abstract:Objective:To investigate the expression of chemokine MIP-3α and its receptor CCR6 in ulcerative colitis(UC) and to explore its relationship with the involvement and severity of UC,so as to asses their expression in the pathogenesis of UC.Methods:The expression of MIP-3α and CCR6 protein in the colon mucosa was detected by immunohistochemistry method in 35 UC patients and 20 normal controls.Results:MIP-3α and CCR6 were both positive in the UC group and negative or only weakly expressed in the normal control group. The expression of MIP-3α and CCR6 in the UC group was significantly higher than that in normal controls (P<0.01). The expression of MIP-3α and CCR6 was related to the severity and involvement of UC. The expression of MIP-3α was significantly correlated with that of CCR6（r=0.765,P＜0.01).Conclusion: The expression of MIP-3α and CCR6 is positively correlated with each other, and both of them participate in the development and progression of UC. The interaction between the 2 may play an important role in the local damage and pathological changes in UC.

Abstract:Objective:To observe the serum level of fibroblast growth factor-23(FGF-23) in patients with end stage renal disease (ESRD) and study its association with phosphorus and vitamin D metabolism.Methods: Serum FGF-23 level was determined by enzyme-linked immunosorbent assay (ELISA) in ESRD patients undergoing haemodialysis (HD, n=50) and peritoneal dialysis (PD, n=24) and in twenty healthy controls (control group,n=20).Serum level of 1，25-(OH) 2VitD was measured by enzyme immunoassay(EIA).Serum intact parathyroid hormone (PTH), creatinine, and calcium and phosphorus were also measured.Results: Serum FGF-23 was obviously higher in HD group（\[88.51±35.01\] ng/L vs \[11.76±3.63\] ng/L）and PD group（\[87.85±33.65\] ng/L vs \[11.76±3.63\] ng/L）than in control group. Moreover, the serum level of 1，25-（OH）2VitD was lower in HD and PD groups than in control group (\[19.82±4.99\] pmol/L vs \[48.37±3.47\] pmol/L; \[24.31±7.11\] pmol/L vs \[48.37±3.47\] pmol/L ), and the level of 1，25-（OH）2VitD was much lower in HD group than in PD group. Pearson relativity analysis showed that serum FGF-23 level was positively correlated with serum creatinine, phosphorus, intact PTH and duration of dialysis(P<0.01); and was negatively correlated with 1，25-（OH）2VitD(P<0.01). Conclusion: Serum FGF-23 is greatly up-regulated in patients with ESRD, and the level of serum 1，25-（OH）2VitD is down-regulated. The changes of FGF-23 is associated with serum creatinine, phosphorus, intact PTH and 1，25-（OH）2VitD.

Abstract:Objective：To investigate the expression of nuclear factor-κB(NF-κB),matrix metalloproteinase-2(MMP-2),tissue inhibitor of metalloproteinase-2(TIMP-2) in the kidneys of rats with acute renal trauma,and to discuss the influence of ulinastatin on their expression and its protective mechanism on the kidney.Methods: The animal model was established by striking the rachi-costaz zone with falling object from the height of 45 cm.Sixty-six rats were randomly divided into three groups:normal control group (C,n=6),trauma group (TRA,n=30),and ulinastatin+trauma (UTI,n=30); the last 2 groups were further divided into 1 h,6 h,12 h,18 h and 24 h subgroups,with 6 animals at each time point.Tissue microarray and immunohistochemistry were used to examine the expression of NF-κB and MMP2/TIMP-2 in different groups.Results: MMP-2 and NF-κB began to express 1 h after trauma in TRA group and their expression was significantly stronger than that in the control group(P<0.05,P<0.01); their expression reached the peaks at 12 h and 6 h after trauma and then gradually decreased.The expression of MMP-2 and NF-κB in UTI group reached their peaks 18 h and 12 h after trauma,respectively,and was significantly higher than that in the control group(P<0.01),but was lower than that in the TRA group at corresponding time points(P<0.01,P<0.05).The expression of TIMP-2 was significantly stronger than that in the control group and TRA group at 6 h,12 h and 18 h after trauma(P<0.01,P<0.05).Conclusion: The expression of NF-κB,MMP-2 is increased in acute

Abstract:Objective:To establish a stress ulcer model by seawater immersion and restriction in rats, and to investigate the mechanism of preconditioning on stress ulcer and the effect of different stressing periods on the ulcer. Methods: A total of 60 male Wistar rats were equally divided into 6 groups , namely, the normal control group (Group A) , direct stress group(Group B), precondition groupⅠ (Group C), precondition groupⅡ(Group S), precondition group Ⅲ (Group E), and ranitidine group (Group F).Precondition peroids of C,D,E groups were 0.5 h/d,1 h/d,and 1 h/d,respectively, for 5 days. Stress time in E group was 10 h,and in B,C,D,and F groups were 8 h. Radioimmunoassay was used to determine the sera levels of endothelin (ET) and 6-dk-dPGF1α in rats. Ulcer index was calculated by Guth method; the ulcer lesion tissues were collected for further pathological examination. Results: The sera ET levels in group C, D and F were lower than those in group B（P<0.01，P<0.05，and P<0.01, respectively）. The sera 6-dk-dPGF1α levels in group C, D and F were significantly higher than those in group B（P<0.01，P<0.05，and P<0.01, respectively）. The ulcer indices in group C, D and F were significantly lower than those in group B（P<0.01，P<0.05，and P<0.01, respectively）. The pathological results were improved in group C, D, and F compared with those in group B. Conclusion: Preconditioning can effectively reduce the rat stress ulcer,but long-time stressing may offset the protective effect of precondition.

Abstract:Objective:To investigate the effects of preventive high thoracic epidural block (PHTEB) for different periods on cerebral vasospasm (CVS) following subarachnoid hemorrhage (SAH) in rabbits.Methods: T6-7 epidural space was opened and an epidural pipe was introduced to establish PHTEB model. Ropivacaine(1 g·L-1) was injected continuously through the pipe at 1 ml/h to block T1-5 sympathetic nerves. Sixty HTEB rabbits were assigned to 6 groups (n=10 each): the normal 1 day group (N1), the normal 7 day group(N7) , SAH 1 day group (S1), SAH 7 day group (S7) , PHTEB plus SAH 1 day group (HS1), and PHTEB plus SAH 7 day group (HS7). CVS was developed by injecting non-anticoagulant autologous arterial blood (0.5 ml/kg) into the cisterna magna. Transcranial Doppler (TCD) ultrasonography was used to determine Vm of the basilar artery. The lesion and spasm of the basilar artery were observed under optical microscope. Results: Compared with group N1, Vm was significantly increased in group S1 (P<0.01) and increased in group HS1 (P<0.05). Compared with group N7, Vm was significantly increased in group S7 (P<0.05) and similar to that in group HS7 (P>0.05). Vm in group S1 was significantly increased compared with group S7 (P<0.05); and that in group HS1 was significantly increased compared with group HS7 (P<0.05). Pathological observation showed no obvious changes in group N1 and N7. The vessel wall was shrunk and the lumina became narrow in group S1 and S7. The pathologic changes in group HS1 and HS7 were slighter than those in group S1 and S7;and Group HS7 was slighter than group HS1. Conclusion: PHTEB can relieve CVS in SAH rabbits, and the relieving effect increases with time of PHTEB.

Abstract:Objective:To observe the changes of serum adiponectin level in patients with non-alcoholic fatty liver disease (NAFLD),and to study the correlation between the serum level of adiponectin and NAFLD. Methods: A total of 110 patients with NAFLD who were hospitalized from Feb. 2006 to Mar. 2008 were included in this study. The patients were divided into non-alcoholic steatohepatitis (NASH) group (n=52,group 1) and simple fatty liver group (n=58,group 2). Fifty-four healthy volunteers were taken as normal control (group 3). The height,weight,and body mass index (BMI) were measured in all groups. The fasting blood samples were obtained to examine serum adiponectin,TC,TG,HDL-C and LDL-C. Stepwise regression method was used to analyze relationship between the serum level of adiponectin and NAFLD. Results: The serum adiponectin and HDL-C levels in group 1 and group 2 were significantly lower than those in group 3 (P＜0.01); the levels of TC,TG,and LDL-C were significantly higher than those of group 3 (P＜0.01). The serum adiponectin in group 2 was significantly lower than that in group 1 (P＜0.05). Lower serum adiponectin and HDL-C levels (P＜0.05) and higher AST level (P＜0.01) were found in group 2 than those in simple fatty liver patients. In non-alcoholic steatohepatitis patients,serum adiponectin level was negatively correlated with the levels of TC,TG (r=-0.436,P＜0.05; r=-0.567,P＜0.01,respectively) and positively correlated with the level of HDL-C (r=0.524,P＜0.01). Conclusion: The serum adiponectin level is correlated with the severity of NAFLD patients,and examination of serum adiponectin may be helpful in understanding the status of NAFLD patients.

Abstract:Objective:To investigate the prevalence and clinical significance of Modic changes in endplates of cervical vertebral body in patients with cervical spondylotic myelopathy(CSM).Methods: The T1-weight and T2-weight sagittal MRI scans of 136 CSM patients undergoing anterior operation were retrospectively reviewed. The patients’ age, gender, prevalence, precise vertebral levels and specific type of Modic changes were recorded, and the association of axial pain with Modic changes was analyzed.Results: Modic changes were observed in 23 patients (16.9%), including 17(17.7%) male and 6(15%) female. The most frequent cervical spinal levels of Modic changes was C5-6. Of all the patients, 4.4％ had type Ⅰ Modic change, 7.4％ had type Ⅱ, and 5.1％ had type Ⅲ. The incidence of axial pain was 56.5% in Modic change groups and 20.4% in non-Modic change groups; there was significant difference between the two groups (P<0.05). Postoperatively, the axial pain was relieved in 76.9% in Modic change groups and 73.9% in non-Modic change groups (P>0.05). The pre-operation incidences of axial pain in patients with type Ⅰ, type Ⅱ and type Ⅲ Modic change were 83.3％, 60％ and 28.6%, respectively; and the post-operation pain-relieving rates were 100％, 66.7％ and 50％, respectively.Conclusion: The most common Modic change is type Ⅱ in the cervical spine, with the C5-6 level being the most frequently involved. The incidence of axial pain is high in patients with Modic change, especially those with type Ⅰ.

Abstract:Objective:To observe the effect of ultrasound ablation combined with the balloon angioplasty in treatment of patients with arteriosclerosis obliteration of lower extremity and diabetic foot.Methods: The 60 limbs of 47 patients with arteriosclerosis obliteration of lower extremity (AOLE) were treated comprehensively by anti-infection,enlarging blood vessel,improving circulation,and anticoagulation; they also received ultrasound ablation,balloon angioplasty and stenting.Results: The patent rate of the 60 limbs was 93.3%.One month and 3 months after treatment with ultrasound ablation combined with balloon angioplasty,the MRI of artery of lower extremity,ultrasound manifestation,ankle-brachial index (ABI),the skin temperature of lower extremity and the sense to coolness,numbness,pain,ochrodermia or cyanosed were all improved to some extents compared with those before treatment.The healing of the diabetic foot was also accelerated.Conclusion: Ultrasound ablation combined with the balloon angioplasty and stenting,together with conventional treatment like anti-infection,anticoagulation,enlarging blood vessel,improving circulation,can reopen the occluded blood vessels and improve blood supply,thus provides a favourable condition for treatment of diabetic foot.

Abstract:Objective:To explore the surgical treatment for well-differentiated thyroid carcinoma（WDTC） invading the larynx,trachea,esophagus and hypopharynx. Methods: Primary tumor resection was performed in 17 patients with advanced WDTC．All the 17 cases had laryngotracheal involvement and received different kinds of surgical treatment: including 14 receiving local shaving-off,wedge tracheal resection with direct suture,window tracheal resection with sternocleidomastoid myoperiosteous flap reconstruction,sleeve tracheal resection with tracheal anastomosis or tracheal reconstruction by forearm free skin flap,and 3 receiving total laryngectomy or near-total laryngectomy. Eleven cases with esophagus and hypopharynx involvement received muscular layer resection or esophageal and hypopharynx partial resection with pectoralis major myocutaneous flap reconstruction. For 12 cases with recurrent laryngeal nerve involvement,9 were treated by nerve decompression,nerve resection combined with one-stage arytenoid within-shift and/or recurrent laryngeal nerve transposition,3 by simple laryngectomy. Results: The patients were followed up for 1 to 12 years. Sixteen patients survived and one died of tumor metastasis. The 5-year survival rate was 93.3%; the post-operation complication rate was 23.5％.Tumor relapses were found in 2 cases who had undergone local shaving-off or partial cervical trachea resection. The voice quality of the 9 patients receiving recurrent laryngeal nerve decompression or voice rehabilitation was greatly improved,and 5 of them had normal voice. Conclusion: The curative rate and quality of life of WDTC patients with extrathyroidal extension can be improved by one-stage resection and reconstruction. Operation modes should be chosen according to the location, involvement of tumor, the invasion of recurrent laryngeal nerve and the disease course. Operation is invasive and reconstruction is a complicated process, with many complications.

Abstract:Objective:To retrospectively analyze the clinical data of 71 patients with upper urinary tract transitional cell carcinoma (UUT-TCC), so as to assess the prognosis of patients and search for factors influencing the prognosis of patients. Methods: Totally 71 UUT-TCC patients treated surgically from Feb. 2004 to Jan. 2007 were included in this study. All patients underwent radical nephroureterectomy via retroperitoneoscopic approach or open approach. The patients were followed up once a month after operation. Traditional prognostic factors including age, sex, tumor stage, grade, location, type of surgical treatment and some pathological biomarkers, such as p53, Ki-67, nm23, and C-erB-2, were collected and analyzed with respect to disease-specific survival by Kaplan-Meier method. Predictive factors influencing the disease-specific survival were also analyzed by Cox proportional hazards model. Results: The 1-, 2- or 3-year disease-specific survival rates were 95.8 %, 89.3% and 83.5%, respectively. Only tumor stage was found to be a factor for prognosis by multivariate analysis (P<0.001). Conclusion: Tumor stage is an independent predictor of disease-specific survival. Retroperitoneoscopic radical nephroureterectomy achieves similar outcomes to the traditional open surgery on oncology control, with the long-term outcome remains to be further studied.

Abstract:Objective:To study the anti-inflammation contents in the stems of Daphne genkwa.Methods: The anti-inflammation contents were obtained from Daphne genkwa by bio-assay guide isolating method. The pharmacological model of dimethylbenzene-induced ear swelling was used for pharmacological study. The fractions of Ligroine and chloroform part of the the EtOH extraction were isolated and purified by column chromatography on silica gel and Sephadex LH-20 and recrystallization. Their structures were studied by using UV，IR,1H-NMR，13C-NMR， and MS, techniques.Results: Nineteen compounds were isolated from the stems of Daphne genkwa and were identified as 2,3-dihydroxypropyl hexadecoate(1), β-sitosterol(2), dueicosanyl caffeate(3), docosyl caffeate(4), octadecyl caffeate(5), daucosterol(6), genkwanin(7), luteolin(8), （+）lariciresinol(9), apigenin(10), kaempferol(11), daphnodorin B(12), genistein(13), dihydrokaempferol(14), p-hydroxybenzonic acid(15), quercetin(16), syringin(17), syringaldehyde(18), ethyl 4-hydroxybenzoate(19).Conclusion: Compounds 1, 3-5, 7, 15, 17-19 have been isolated from the stems of Daphne genkwa for the first time. The high dosage of petroleum ether extract, chloroform extract and compound 4 have significant anti-inflammation activity in mice with ear swelling.

Abstract:Objective:To prepare fluorouracil-loaded chitosan nanoparticle (5-Fu-CS-NP) for ophthalmologic usage and evaluate its releasing characteristics in vitro. Methods: 5-Fu-CS-NP was prepared with 5-Fu, polyacrylic acid and chitosan using dispersion method. The mean size, entrapment efficiency (EE%), drug loading (DL%) and the in vitro releasing characteristics of 5-Fu-CS-NP were investigated.Results and conclusion: The mean size of 5-Fu-CS-NP was (144.6±3.1) nm, the EE% was 84.5% and the DL% was 3.91%. 5-Fu-CS-NP had a satisfactory sustained-releasing effect and the releasing could last for 3-7 days, and the release behavior was not greatly changed within a pH value of 7.2-7.4.

Abstract:Objective:To construct lentivirus carrying both novel human pancreatic cancer gene S100P and green fluorescent protein (GFP) gene.Methods: The fragments containing all the exons of S100P were amplified by RT-PCR and were cloned into the lentivirus vectors labeled with GFP. The lentivirus was packaged and was used to transfect 293T cells together with pShuttle. The supernatant of virus-producing cells was harvested, concentrated, identified, and was used to infect 293T cells and pancreatic cancer cells. Fluorescent microscopy was used to observe the fluorescence in the 293T cells; and real time-PCR was used to examine the relative contents of S100P in pancreatic cancer cells.Results: Electrophoresis showed that the sequence of the RT-PCR product was consistent with the data of NCBI by DNA sequencing analysis. The lentivirus effectively transfected 293T cells. Strong green fluorescence was observed by fluorescent microscopy. The supernatant of lentivirus-transfected 293T cells effectively infected 293T cells and the relative content of S100P in the transfected pancreatic cancer cells was higher than that of control group.Conclusion: The lentivirus vector containing S100P-GFP recombinant gene have been successfully constructed, which provides a basis for further study of S100P function.

Abstract:Objective:To exploit marine microorganisms and study their secondary metabolites for new drugs.Methods: An antibacterial model was used to screen for active strains. The ethyl acetate (EtOAc) extract was separated by silica chromatography, gel filtration chromatography, and high-performance liquid chromatography. The structures of the compounds were elucidated by 1HNMR, 13CNMR and MS technologies; Kirby-Bauer Disc Diffusion method and MTT method were employed to detect the biological activities of the separated compounds.Results: Eleven compounds were separated and identified as macrolactin A (1), 3-Hydroxyl acetyl-indole (2), 3-indolethanol (3), cyclo-(Try-Pro) (4), cyclo-(Ile-Try) (5), cyclo-(Leu-Pro) (6), cyclo-(Leu-Val) (7), cyclo-(Ile-Pro) (8), cyclo-(Phe-Val) (9), N- phenethylacetamide (10), P-hydroxy benzaldehyde (PHB) (11).Conclusion: Compound 1 shows strong inhibitory activities against Pyricularia oryzae, Escherichia coli and Staphylococcus aureus (with MIC values being 3.6, 0.45 and 6.3 μg/ml, respectively), and tumor cell lines HeLa and HepG2 (with the IC50 values being 2.0 and 1.8 μg/ml, respectively).

Abstract:Toumor microenvironment is closely related to the development and progression of tumors.Recently, researchers suggest that abnormal differentiation of stem cells might lead to the initiation of cancer.Microenvironment is the key to homeostatic regulation of stem cells.Disregulation of microenvironment plays an important role in the carcinogenesis.And during the development of tumor,a suitable microenvironment can promote the rapid proliferation of tumor.Changing the specific microenvironment can suppress the tumorigenic phenotype of aggressive cancer.Meanwhile,microenvironment also plays an essential role in cancer metastasis.Understanding of these underlying mechanisms will enhance our knowledge of cancer and will provide a novel basis for prevention and treatment of cancer..

Abstract:Currently peripherally inserted central catheter (PICC) is a commonly used venous catheter technology in clinical background. It has been controversial as to where the end of PICC should be placed and how to decide its position under X-ray. This paper reviews the latest studies on PICC, including the optimum location, imaging markers for positioning under X-ray, and the mobility of the end of PICC.

Abstract:外伤性面神经损伤导致的面瘫等临床表现对患者的生活、工作带来诸多的不良影响。颅脑创伤和医源性损伤是造成面神经损伤的主要因素,外伤性面神经损伤研究是国内外学者研究的热点之一,热点内容主要集中在以下几个方面：面神经损伤修复与再生微环境的营造，医源性面神经损伤的减少，临床检查手段的完善，手术治疗的时机，指征、入路和方法规范等。本文就这些热点问题予以讨论。

Abstract:目的:研究团体心理咨询对军事应激状态下军人应对方式和状态-特质焦虑的影响，寻找调控军事应激的有效方法。方法:团体心理咨询前1 d(前测)、咨询后1 d(即时后测)以及咨询后1个月(长效后测)，采用简易应对方式问卷(SCSQ)、状态-特质焦虑问卷(STAI)和自编一般情况调查表对204名即将参加军事演习的军人进行集体心理测试。结果: 前测时积极应对因子和消极应对因子平均分为(25.31±3.57)和(13.76±3.61)；特质焦虑与状态焦虑平均值分别为(38.20±6.41)和(42.21±7.17)。不同文化程度和职别的军人应对方式差别显著。状态-特质焦虑与应对方式存在相关性。 即时后测时积极应对因子和消极应对因子平均分为(28.73±3.49)和(10.46±3.65)；特质焦虑与状态焦虑平均值分别为(37.98±6.94)和(38.16±6.43)。长效后测时积极应对因子和消极应对因子平均分为(27.96±3.85)和(11.02±3.42)；特质焦虑与状态焦虑平均值分别为(37.45±6.37)和(38.13±7.02)。咨询前后积极应对因子、消极应对因子、状态焦虑平均分差异有统计学意义 (均为P<0.001)。结论:团体心理咨询能有效地提高军事应激状态下军人的应对能力，减轻状态焦虑，是调控军事应激的有效干预措施。

Abstract:目的:比较两种腹腔镜入路对泌尿系外科手术的影响。方法:选择腹腔镜下行泌尿外科手术的患者21例，其中经腹腔入路(经腹组)12例，经腹膜后入路(腹膜后组)9例，记录麻醉后充气前、充气后、气腹解除后血压(BP)、心率(HR)、平均动脉压(MAP)、呼气末二氧化碳(PETCO2 )、气道峰值压力(Ppeak)、分钟通气量(MV)和动脉血气(ABG)；麻醉后充气前、充气后血糖(Glu)、血皮质醇(CORT)和促肾上腺皮质激素(ACTH)。结果:两组充气后Ppeak、MV、PETCO2、Gul、CORT和ACTH水平明显升高，但组间无统计学意义。结论:两种腹腔镜充气方式下呼吸、循环和应激反应的变化过程基本相似。

Abstract:目的:观察幽门螺杆菌(Hp)所致不同疾病中，血清与胃液内非对称性二甲基精氨酸(ADMA)与一氧化氮(NO)的浓度变化，初步探讨ADMA在Hp所致不同疾病中的作用。方法:139例患者，分为Hp阳性组(n=72)，其中胃炎24例，胃溃疡13例，十二指肠球部溃疡35例;男42例，女30例，年龄20～50岁，平均(33.6±10.4)岁；Hp阴性组(n=67)，其中胃炎47例，胃溃疡6例，十二指肠球部溃疡14例，男40例，女27例，年龄19～54岁，平均年龄(34.9±11.5)岁。检测两组患者血清与胃液内ADMA与NO浓度并进行组内与组间对比。结果:胃液内ADMA和NO浓度Hp阳性组明显高于Hp阴性组(P<0.05)，血清内ADMA和NO浓度在两组患者之间无明显差别。Hp阳性患者不同病变之间，胃炎患者胃液内ADMA浓度最高，胃溃疡患者次之，十二指肠溃疡患者最少(P<0.05)；而NO浓度以十二指肠溃疡最高，胃溃疡次之，胃炎患者最少(P<0.01)。结论:Hp感染导致患者胃内ADMA与NO浓度增加，且二者在不同病变中变化程度不同，说明Hp的致病作用可能与ADMA调节NO的产生有关。

Abstract:目的:探讨肝内胆管结石合并肝胆管癌的早期诊断和治疗措施。方法:回顾性分析近6年收治的18例肝内胆管结石并发肝胆管癌患者的临床资料。结果:本组肝胆管结石并肝内胆管癌的发生率为5.0% (18/359),癌肿均发生于含结石胆管处,术前B超、CT、MRCP/ERCP对肝胆管癌变的诊断正确率分别为57.1% (4/7)、70% (7/10)、45.5% (5/11);手术切除率为61.1%(11/18),根治性切除率27.8% (5/18)。术后1、3年生存率:根治性切除组分别为80% (4/5)、40% (2/5)，姑息性切除组分别为33.3% (2/6)、16.7% (1/6),两者比较差异具有统计学意义(P<0.01)。结论:肝内胆管结石长期、反复发作是引起胆管癌的重要原因;肝胆管结石癌变的延误诊断率高、根治性切除率低、预后差。早期诊断并手术切除依然是肝胆管结石癌变最有效的治疗方法。

Abstract:目的:比较采用动力髋螺钉(dynamic hip screw，DHS)、Gamma钉和防旋型股骨近端髓内钉(proximal femoral nail anti-rotation,PFNA)治疗老年骨质疏松性股骨粗隆间骨折的效果，探讨老年骨质疏松性股骨粗隆间骨折合理的内固定治疗方法。方法:2000年2月至2006年10月，分别采用DHS(DHS组，n=59)、Gamma钉(Gamma钉组，n=46)和PFNA(PFNA组，n=53)治疗并随访老年骨质疏松性股骨粗隆间骨折患者158例。比较3组患者手术情况、术中和术后并发症、疗效及骨折愈合时间。结果:各组间平均手术时间和术中出血量两两比较均有统计学差异(P＜0.01)。三组术中、术后并发症发生率有统计学差异(P＜0.05)，PFNA组术中、术后并发症发生率比DHS组和Gamma钉组低。DHS组、Gamma钉组和PFNA 组优良率分别为86.44%、84.78% 和90.57%，组间疗效差异无统计学意义(P>0.05)。PFNA组与DHS组骨折愈合时间有统计学差异(P＜0.01)，而Gamma钉组与PFNA组间无统计学差异(P>0.05)。结论:应用DHS、Gamma钉和PFNA治疗老年股骨粗隆间骨折，在疗效方面无明显差异。 PFNA内固定平均手术时间最短，术中出血量最少,术中、术后并发症发生率低,骨折愈合时间短。PFNA对骨折端的血循环影响和骨质破坏较小,固定更为牢靠,是治疗老年骨质疏松性股骨粗隆间骨折的合理手术方法。

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Abstract:1临床资料患者，女，29岁。2年前无明显诱因出现腰酸，以左侧为著，无血尿、腰痛、发热等症状。X线、CT检查提示“左侧完全性重复肾”，于2007年7月20日入院拟行重复肾切除术。体格检查无异常。超声检查见左肾形态饱满，左肾集合系统分隔成互不相通的两部分，并有各自相连的输尿管。上位肾集合系统分离，最宽处约60 mm，液体内透声良好；上位输尿管全程扩张，内径约12 mm，其末端呈一大小约38 mm×24 mm的囊肿突入膀胱腔内，有裂隙样开口与膀胱腔相通；下位集合系统及输尿管未见扩张。右肾大小形态正常，包膜光整，皮髓质分界清，集合系统分为互不相通的上下两部分，并有各自相连的输尿管，上位集合系统分离约8 mm，但上位输尿管以及下位集合系统均未见明显扩张。

Abstract:胡桃夹综合征(nutcracker syndrome,NCS)又称左肾静脉压迫综合征,是由于先天或后天形体变化等原因致使腹主动脉和肠系膜上动脉之间夹角变小,左肾静脉受到两者的挤压导致肾静脉血回流受阻,从而引起的以血尿、蛋白尿为主的泌尿系综合征\[1\]。精索静脉曲张是导致男性不育的一个主要原因,文献报道在左侧精索静脉曲张的患者中有72%伴有左肾静脉受压\[2\]。我们采用开放式精索静脉高位结扎术治疗了8例NCS合并左侧精索静脉曲张的患者,现报告如下。

Abstract:1临床资料女性患者，56岁，3年前因右肾区不适诊断为肾癌行右全肾切除，半年前无意中发现右腮腺无痛肿块，无症状，不影响日常生活。 大体标本检查：圆形肿物，附少许涎腺组织，直径2 cm，包膜完整，与周围组织界限清楚，切面棕黄色，质地软。光镜检查：肿瘤呈浸润性生长，肿瘤细胞呈透明状，实性巢状、条索状排列，部分区呈管状、腺泡状等，细胞柱状、立方状，胞质透明，核圆形或椭圆形，染色质细腻，可见核仁,间质血管丰富，纤维少(图1)。复习肾癌组织H-E切片，图像和腮腺区肿物相同，细胞核圆形，大小比较一致，染色质细腻，可见单个核仁，肿瘤包膜、肾脏被膜及肾周脂肪无浸润，间质未见明显炎细胞浸润。患者家族中无肾癌患者。超声和影像学检查未发现其他转移灶。腮腺区肿物免疫组织化学标记及结果如下：CK(+)，CD10(+)(图2),Vimentin(+)，EMA(+)，CK8(+)，CK18(+)，CK19(+)，S-100(-)，GFAP(-)，Actin(-)，Myosin(-)， Calponin(-)， SMA(-)，CK14(-)，HCK(-)。病理诊断：(右腮腺)结合病史，肾脏透明细胞癌腮腺转移。

Abstract:头孢菌素类抗生素具有抗菌谱广、杀菌作用强、毒性较低、过敏反应较少等优点,而广泛应用于临床。但随着临床应用增多其不良反应的病例日趋增多,轻者给患者增加不适或造成各重要脏器的可逆性损伤,严重者可发生过敏性休克,抢救不及时或引起严重的并发症常可致死,严重影响患者的生命安全。为使其更安全有效服务于患者,本文就1例头孢噻肟钠致高热、血细胞严重降低和肝功能损害报告如下。