Objective To study the sub chronic toxicity of silver nanoparticles on medaka.Methods Adult fish were divided into silve nenopartides and control group.Animals were collected on 14 days after exposure, and some toxicological endpoints such as death rate, tissue distributed of silver irons, oxidative stress and histopathological damage were measured.Results There were significant difference in death rate of medaka treated with silver nanoparticles and control group.Compared with the control group, the content of silver in gill, intestine and liver of medaka treated with silver nanoparticles were increased significantly.Compared with control group, the activity of LDH in liver and SOD in liver and gill were significantly decreased(P<0.01).The content of MDA in liver of medaka treated with silver nanoparticles was significantly increased(P<0.01).The liver and gill of mekada treated with silver nanoparticles were damaged, compared with control group.Conclusion Nano silver has a certain subchronic toxicity to aquatic life.

Objective To study the early diagnosis and management of the patients with acute respiratory distress syndrome (ARDS) caused by cytomegalovirus (CMV) pneumonia after liver transplantation.Methods The clinical data of 8 patients with ARDS caused by CMV pneumonia after liver transplantation in our hospital from April 2001 to May 2004 was retrospectively analyzed. All cases were treated with intravenous infusion of gancyclovir, reduced dosage of cyclosporine A or tacrolimus to 1/3~1/2 of baseline and withdrawal of MMF and prednisone. The patients were subjected to breathing machine assist ventilation and nutrition supply.Results Five patients recovered and 3 died. No one developed acute rejection. Conclusions The key of early diagnosis lies in combining chest X-ray or CT scan with clinical presentation. Administration with anti-viral drugs, adjustment of immunosuppressive agents, management with breathing machine assist ventilation and effective nutrition supply are important for the treatment of patients with ARDS caused by CMV pneumonia after liver transplantation.

Objective To determine the optimal cut-off value of critical-care pain observation tool (CPOT) in assessing degree of pain in patients undergone craniotomy, and to determine the sensitivity and specificity of CPOT with this cut-off value. Methods A prospective observational study was conducted in Beijing Tiantan Hospital. A total of 118 patients admitted to intensive care unit (ICU) after craniotomy was consecutively enrolled during August 2014 to August 2015. CPOT and visual analogue scale (VAS) were used to assess the pain before, during and 20 minutes after the removal of central venous catheters, and the difference was compared between two scores at three time points. Receiver operating characteristic (ROC) curve was used to determine the optimal cut-off values for evaluation of the sensitivity and specificity of CPOT. Patients' complaint of pain was considered the gold-standard. Results CPOT values (inter-quartile range) before, during and after the procedure were 0 (0-3), 0 (0-6) and 0 (0-2), respectively; while VAS values were 4 (1, 6), 3 (1, 6) and 4 (1, 6), respectively. CPOT value during the procedure was significantly higher than CPOT values before and after the procedure (both P < 0.01). When the optimal cut-off value of CPOT was 1, CPOT showed the highest Youden index before, during and after the procedure (1.183, 1.515, and 1.438, respectively), and showed high specificity (all 100%) and low sensitivity (18.3% and 43.8%, respectively) when assessing the pain before and after the removal of the catheter. The sensitivity and the specificity were high when assessing the pain during the procedure, the sensitivity was 69.4%, and the specificity was 82.1%. When the optimal cut-off value of VAS was 2 before and during the procedure, and was 4 after the procedure, VAS showed the highest Youden index, 1.568, 1.452, and 1.509, respectively. VAS demonstrated high sensitivity and specificity before, during and after the procedure (sensitivity was 97.2%, 95.2% and 75.0%, respectively; specificity was 59.6%, 50.0% and 75.9%, respectively). The area under ROC curve (AUC) of CPOT before, during and after the procedure were 0.592 [95% confidence interval (95%CI) = 0.490-0.693], 0.778 (95%CI= 0.693-0.863) and 0.719 (95%CI = 0.627-0.811), respectively; the AUC of VAS before, during and after the procedure were 0.846 (95%CI = 0.771-0.920), 0.767 (95%CI = 0.681-0.854) and 0.838 (95%CI = 0.767-0.909), respectively. The AUC of VAS before and after the procedure was significantly higher than the AUC of CPOT (P < 0.001 and P = 0.006), while there was no significant difference between the AUC of VAS and CPOT during the procedure (P = 0.826). Conclusion CPOT can be used to assess the pain during painful procedure, and it shows high accuracy, but with poor evaluation effect on pain in rest.

BACKGROUND:The hyperpoladzation-activated cyclic nucleotide-gated cation channel (HCN) gene is not increase the risk of induced cardiac arrhythmia,and can accept the modulation function of autonomic nervous system.Therefore,it is the first candidate gene for biological pacemakers.OBJECTIVE:To construct recombinant adenovirus vector containing human HCN4 gene and evaluate its transfection efficency in rat bone marrow mesenchymal stem cells (MSCs).DESIGN,TIME AND SETTING:The in vitro cytology-gene experiment was performed at the Lin Bai-xin Experimental Center,Second Hospital of Sun Yat-sen University from February to September 2008.MATERIALS:Ten SD rats were supplied by the experimental animal center of Sun Yat-sen University.Rlasmid pcDNA3.1-HCN4 containing target gene human HCN4,human embryo kidney 293 cells and Escherichia coll DH5α were preserved by our experimental center;Shuttle plasmid pShuttle-CMV containing green fluorescent protein gene and adenoviral backbone plasmid pAdxsi were bought from SinoGenoMax Co.,Ltd.METHODS:HCN4 cDNA segment was liberated from the cloning vector of pcDNA3.1-HCN4 via Hind Ⅲ+Xba Ⅰ,and subcloned into pShuttle-CMV,which was digested by I-Ceu Ⅰ +I-Sce Ⅰ double enzyme and subcloned into adenoviral plasmid to form recombinant adenovirus plasmid.Recombinant adenovirus plasmid was transfected into 293 cell lines by liposome,and the recombinant adenovirus AdHCN4 was packaged and transfected into rat MSCs.MAIN OUTCOME MEASURES:The identification of recombinant adenovirus plasmid vector,identification of recombinant adenovirus and its titration test;the transfection efficiency of recombinant adenovirus.RESULTS:Cloned sequence about 3.6kbp was obtained by Hind Ⅲ+Xho Ⅰ digestion after HCN4 cDNA segment was cloned into pShuttle-CMV.DNA sequencing results indicated that the clone location was correct.Recombinant adenovirus plasmid was cut into seven fragments while empty vector gained only six fragments after digested by Xho Ⅰ.The recombinant adenovirus was pathogenic to 293 cells after recombinant adenovirus plasmid was packaged in it.HCN4 cDNA (657bp) was amplified by PCR with virus titer of 2.5×10~(11) PFU/mL after transfected 293 cells with supematant.The efficiency of recombinant adenovirus infecting rat MSCs was about 90% when multiplicity of infection was 800.Rat MSCs expressed green fluorescence after transfection.CONCLUSION:The adenovirus vector with human HCN4 cDNA was established successfully,which can effectively transfect rat MSCs.

Objective To evaluate the safety and feasibility of laparoscopic hepatectomy for hepatocellular carcinoma(HCC). Methods From January 2002 through December 2007,86 HCC cases were divided into laparoscopic hepatectomy group and open hepatectomy group.Clinical data were analyzed. Results There were 36 cases in LH group,and 50 cases in OH group.Significant differences were noticed in the length of incision,operative blood loss,food-taking time,postoperative hospital stays,and analgesic usage between the two groups (respectively t =-37.608、-2.396、-13.073、-4.283 、x2 = 35.765,all P＜0.05),in which LH group was superior to OH group.Differences appeared in ALT,AST,ALP,r-GT and LDH on post-op day 1,and 3.APTT,ALB response on the fifth day after operation were different in the two groups (separately t =-3.465,-3.236,-3.470,-6.812,-4.837 and-3.998,-2.894,-4.286,-7.887,-5.388,6.131,7.292; all P ＜0.05);ALT,AST,ALP,r-GT value on the fifth day post-operation,and T-BIL on the day 1,5 post-operation were different in two groups (separately t =-4.795,-2.155,-3.442,-4.194,-2.712,-1.600,all P ＜0.05),Meanwhile,as the results all showed that,injuries were all less severe in LH than OH group.There were no significant differences between the two groups in operative time,resection method,overall complications,1-year,3-year survival rate,disease free survival (t =-0.893,separately x2 =0.066,0.026,0.468,0.156,0.106,2.732,all P ＞0.05) while 3-year survival rate in LH group and OH group were 50.0%,25% respectively (x2=2.732,P = 0.098). Conclusion Laparoscopic hepatectomy was safe and feasible for treatment of HCC,and its short-term efficacy was better than open hepatectomy.Furthermore,laparoscopic hepatectomy had promising long-term effectiveness.

Purpose To study the clinicopathologic features and differential diagnosis of primary mucosal melanoma of the nasal cavity (PMMNC).Methods 17 cases of PMMNC diagnosed from January 2003 to September 2016 were studied by clinical pathological analysis and immunohistochemical staining,and relevant literatures were reviewed.Results 73％ of the PMMNC was characterized by unilateral nasal congestion and intermittent epistaxis and 61％ of the PMMNC occurred in the nasal septum and nasal side wall.Microscopically,the organizational structure and morphology were complex and diverse,which had several cell types including epithelioid cell type (6cases,35.3％),spindle cell type (3 cases,17.6％) and snall cell type (5 cases,29.4％),the other 3 cases (17.6％)were mixed cell type.Mitotic activity and tumor necrosis were more likely to be seen in PMMNC,among other clinicopathological features with a small amount of fibrous stroma and melanoma and rich blood vessels.The immunohistochemical study showed that the positive rate of S-100 and HMB-45 were both 93.8％(15 cases) and those of Melan-A and vimentin were both 87.5％ (14 cases),while CK and EMA were both negative (16 cases).Conclusion PMMNC is a rare disease and the phenotype of S-100,HMB-45,Melan-A and vimentin are useful for diagnosis of PMMNC.

OBJECTIVE:To optimize the extraction technology of Compound qima capsules. METHODS:With the blood pres-sure lowering of rats as index,pharmacological efficacy test was used to screen the preparation technology(A was whole herb de-coction;B was Gastrodia elata fine powder mixed with other decocted medical materials). The extraction technology was opti-mized by single factor and orthogonal test using the contents of astragaloside and isoflavone grape glycosides and the quality of sol-id as indexes,with added water,decoction time,decoction times as factors;and the verification test was carried out. RESULTS:Pharmacological efficacy test showed that antihypertensive effect of sample by technology B was superior. The optimal extraction condition of other medical materials of technology B was as follows as 12-fold water per time,decocting for 1.5 h,for 3 times. In verification test,average extraction rates of astragaloside and isoflavone grape glycosides were 64.02% and 51.97%,and average value of the quality of solid was 5.69 g(RSD≤1.92%,n=3). CONCLUSIONS:The optimized extraction technology is stable and feasible.

AIM: To investigate the influence of Ginkgo biloba extract (EGb761) on c-jun expressions and motoneurons survival following root avulsion. METHODS: One hundred and eighty adult Sprague-Dawley female rats were randomly divided into control and EGb761 groups. Immediately after avulsion of C5-T1 nerve roots, the rats were injected ip with either 1 mL of EGb761 25 mg?kg~(-1)?d~(-1) or the same volume of normal saline, and the treatment repeated everyday. At 4 h to 6 weeks following avulsion, the C7 spinal segments of all rats were collected and prepared for c-jun immunocytochemistry and neutral red stain. The numbers of (c-jun) positive and survival motoneurons were counted and compared between two groups at each time point. RESULTS: In control rats following avulsion, c-jun positive motoneurons appeared at 4 h, reached its maximum at 1 d and declined to 2 weeks. Avulsion-induced motoneurons death started at 2 weeks, climbed to its maximum at 4 weeks-6 weeks. In EGb761 treated rats, both numbers of c-jun positive and survival motoneurons were more than that in control group at each time point. CONCLUSION: EGb761 attenuates avulsion-induced motoneurons death, and this effect may be related to up-regulation of c-jun gene in avulsed motoneurons. [

Objective To develop neural stem cells(NSCs) which can stably express exogenous brain-derived neurotrophic factor(BDNF) in vitro. Methods NSCs from the subependymal zone of embry-onic day 14.5(E14.5) rat brain were purified by limiting dilution assay and then infected with supernatant of recombinant retrovirus pLXSN-BDNF and retrovirus pLXSN. The original copy numbers of exogenous gene templates from three groups NSCs(pLXSN-BDNF viral infection group, pLXSN viral infection group, control group) were detected by fluorescent quantitative PCR(FQ-PCR). ELISA assay was used for determining the protein contents of BDNF of supernatant from three groups NSCs for six days continually after seeded in 24-well plates in the same cell density. Results NSCs were purified successfully by limiting dilution assay.The original copy numbers of exogenous BDNF gene templates from pLXSN-BDNF viral infection group by FQ-PCR were (19.57±0.65) × 10~3 copies/μl, higher than those of another two groups(P < 0.05). The protein contents of BDNF of supernatant from NSCs of pLXSN-BDNF viral infection group was highest among three groups and compared with another two groups had statistical significance (P <0.05) . Conclusion The purified NSCs can be transduced exogenous BDNF successfully with supematant of recombinant retrovir-us pLXSN-BDNF which provide experimental evidences and laying foundations for further research of retinal transplantation and quantization investigation of gene therapy for optic nerve injury.

Objective To explore the change of serum osteopontin levels in preoperative and postoperative patients with non-small cell lung cancer (NSCLC) and its clinic significance.Methods The levels of serum osteopontin were detected by ELISA.Results In 88 NSCLC patients,serum osteopontin levels were significantly higher than in benign pulmonary disease and healthy control group(P＜0.05.P＜0.01.respectively) There was no difference between benign pulmonary disease and healthy control group(P＞0.05).Serum osteopontin levels were related to clinical stage and lymph node metastasis,and the levels of serum osteopontin were lower in postoperative patients than in preoperative cases(P＜0.05).For NSCLC.the detection of osteopontin had a sensitivity of 84.1%,a specificity of 92.5%,and an accuracy of 88.1%. Conclusion Increased serum levels of osteopontin may be useful as a helpful clinical biomarker to diagnose earlier,predict prognosis and differentiate from benign pulmonary disease.