BACKGROUND: Monoamine hypothesis has been demonstrated by researches. However, the correlation between the metabolite of plasma monoamine neurotransmitter and anti-depression treatment in patients with depression has less been reported.OBJECTIVE: To study the effect of different drugs on metabolite of plaama monoamine neurotransmitter, and the correlation between the metabolite of plasma monoamine neurotransmitter and anti-depression treatment in patients with depression.DESIGN: Case controlled study.SETTING: Neurological Department and Brain Institute of Nanjing Medical University.PARTICIPANTS: Forty patients with depression hospitalized in Nanjing Brain Hospital (depression group) were diagnosed with the second revised edition of China classification of mental diseases(CCMD-2) and the tenth edition of International classification of diseases. And the total score of Hanmilton rating scale for depression(HAMD) was more than 17. Healthy voluntary blood donators in the control group were from Nanjing Municipal Central Blood Station( n = 20).INTERVENTIONS: Antidepressant was used in the depression for 4 weeks: fluoxetine 20 mg per day; 5-serotonin selective reuptake inhibitor (SSRI) paroxetine 20 mg per day; venlafaxime 50- 100 mg per day;5-serotonin and morepinephrine selective reuptake inhibitor(SNRI) fluvoxamine 50-100 mg per day. High performance liquid chromatograpy(HPLC)was used to measure the level of metabolite of plasma monoamine neurotransmitter in patients with depression before and 42 week after treatment, and the HAMD was used to evaluate clinical effect of the patients.MAIN OUTCOME MEASURES: The levels of metabolites of plasma monoamine neurotransmitters in patients with depression: 5-hydroxyindoleace tic acid(5-HIAA), 3-methoxy-4-hydroxyphenylglycol(MHPG) and homovani llic acid(HVA) were measured before and 4th week after treatment.RESULTS: The levels of 5-HIAA, MHPG and HVA of the metabolites of plasma monoamine neurotransmitters in patients with depression before treatment [ (20.3±14.6), (124.8±103.6), (54.7±32.1) μg/L] were all lower than those in the normal control group[ (39.5±28.4), (334.5 ±107.3), (88.5±37.2) μg/L], with statistically significant differences (P＜0.05). After SSRI treatment, the 5-HIAA content[ (37.1±21.9)μg/L]was significantly increased as compared with that before treatment, whose difference indicated significant meaning ( P＜0.05), but the differences in MHPG and HVA had no significant meaning as compared with those before treatment(P＞0.05) . After SNRI treatment, 5-HIAA and MHPG contents [(35.4±25.2 ), (291.2±120.4) μg/L] both were significantly increased, which indicated significant difference as compared with those before treatment( P＜0.05); but HVA level had no significant changes.CONCLUSION:'The peripheral neurotransmitter metabolites in plasma can reflect their states in brain. The change of neurotransmitter metabolite in plasma can be regarded as an important reference index for the evaluation of depression.

The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.

OBJECTIVE To investigate the hospital infection rate and sites of patients in emergency intensive care(unit)(EICU),and provide basis for prevention and treatment of hospital infection.METHODS Patients who were hospitalized in EICU were investigated by retrospective study.RESULTS Among the hospital infection sites,(respiratory) tract was the most frequent one(65.63%),the next was urinary tract(28.13%),and the deep vein was the third(6.25%).The most common hospital infection bacteria were Staphylococcus aureus,Pseudomonas aeruginosa and Acinetobacter baumannii.CONCLUSIONS The analysis of the subjective and objective factors of hospital infection,and the acknowledge of relationship between nursing and hospital infection sites are(important) basis for hospital infection prevention and treatment.

Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.

· AIM :To investigate the role of pericytes in growth of retinal microvascular endothelial cells with a co-culture system in order to understand some mechanism of angiogenesis in hypoxia induced retinal neovascular disorders.(RMECs) were isolated by a modified protocol using CD31 coated Dynabeads, and identified by immunocytochemical staining with anti-Factor Ⅷ and CD31 antibodies. Rat retinal pericytes were isolated and characterized by immunofluorescent staining with PDGFR-β; and desmin antibodies. Pericytes and RMECs were cultured in a contact co-culture system both under normoxia and hypoxia by Millicell chamber. RMECs proliferation was evaluated by MTT and cell cycle assay with flow cytometry. RT-PCR was used to detect the alteration of KDR/Flk-1 mRNA level in RMECs under normoxia or hypoxia in the co-culture system.harvested with the modified isolating method. The two cell types were identified by positive Factor Ⅷ, CD31 and PDGFR-β, desmin cytochemical staining respectively.RMECs proliferated significantly under hypoxia from 3 to 9d with a maximal rate on day 6 (24.9%, P ＜ 0.01) by MTT. In the co-culture system, the proliferation of RMECs was inhibited by pericytes. After 6d exposure to hypoxia,the fraction of S-phase RMECs number was greatly increased by 43.9% (P ＜ 0.01). In the co-culture system,RMECs proliferation was inhibited by pericytes through decreasing the fraction of S-phase cell number both under normoxia (3.6%, P＜0.05) and under hypoxia (15.1%,P＜0.01). KDR/Flk-1 mRNA level in single cultured RMECs was shown to increase approximately 1.3-fold when exposed to hypoxia. Compared with single cultured RMECs, co-culture with pericytes could decrease KDR/Flk-1 mRNA by 45.1% (P＜0.05) and 27.7% (P ＜ 0.05) under normoxia and hypoxia condition respectively.pericytes could inhibit proliferation of RMECs under both normoxia and hypoxia. The inhibition effects of pericytes maybe, at least in part, due to downregulation of KDR/Flk-1 of RMECs. These findings confirm that pericytes could be a potential inhibitor in the pathogenesis of retinal neovascularization.

OBJECTIVETo study the killing effects of the liposome-mediated thymidine kinase (TK)/ganciclovir (GCV) system on MG-63 osteosarcoma (OS) cells and its bystander effects.

METHODSLiposome-mediated TK gene transfected into MG-63 OS cells, the efficiency of transfection was analyzed by flow cytometry and observed under inverted fluorescence microscope. Non-transfected osteosarcoma MG-63 cells were divided into three groups,in the experimental group 1 transfected TK/GCV cells cultured in solutiona liquid mixture by supernatant by 1/10,1/7,1/5,1/2 ratio to original broth; in the experimental group 2 transfected cells cultured in solutiona liquid mixture of supernatant filtered through 0.22 microm filter by 1/10,1/7, 1/5, 1/2 ratio to original broth, in control group the transfection cells cultured in original culture solution. Cell growth inhibition rate and osteosarcoma cell sensitivity to TK/GCV system were measured by MTT assay in each group.

RESULTSThe TK gene was transfected into MG-63 OS cells successfully by liposome-mediated, flow cytometry instrument detection TK gene transfection cell transfection efficiency can reach 75.5%. Six days later the MTT assay showed that in the experimental group 1 inhibition rate of all concentration ratio of the mixed culture fluid were statistically significant as compared with the control group (P < 0.05), and in the experimental group 2 that of the 1/10 and 1/7 of concentration ratio of mixed culture medium was not statistically significant as compared with the control group (P > 0.05). TK gene transfected MG-63 cells increased with the the GCV concentration,the cell apoptosis rate increased.

CONCLUSIONThe experiment demonstrated that the MG-63 OS cells are sensitive to the liposome-mediated TK/GCV system and bystander effects are significant.

Apoptosis ; drug effects ; Bone Neoplasms ; enzymology ; genetics ; physiopathology ; Bystander Effect ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Ganciclovir ; toxicity ; Humans ; Osteosarcoma ; enzymology ; genetics ; physiopathology ; Thymidine Kinase ; genetics ; metabolism ; toxicity

Objective To compare the perioperative outcomes of laparoscopic liver resection (LLR) versus open liver resection (OLR) for hepatocellular carcinoma (HCC).Methods A total of 89 HCC patients undergoing liver resection between January 2012 and November 2016 were enrolled.Nonparametric tests were employed to compare the clinicalpathological characters and preoperative outcomes.Results No significant difference was observed in clinicalpathological features and postoperative morbidity.LLR group had shorter hospital stay (Z =4.642,P ＜0.01),lower serum ALT level in 1st,3rd and 5 day (Z =2.157,3.089,2.384,all P ＜0.05) and AST level in 1st-and 3rd-day postoperatively (Z =2.688,2.566,all P ＜0.05).The growth rate in serum total protein (TP) and albumin (ALB) postoperatively is higher for LLR group (y =2.348 4x + 51.696 vs.y =0.902 9 + 35.532),(y =1.539 9x + 29.68 vs.y =0.732 9x + 30.406).Conclusion LLR allows quicker liver function recovery and shortens patients' postoperative hospital stay.

In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.

Long-term excessive iodine intake resulted in an increased TT_4 level and a decreased TT_3 level in maternal serum,meanwhile,hepatic and renal type 1 deiodinase activity decreased dose-dependently.A significant reduction in type 2 deiodinase ( D2 ) activity of 12.5 d placenta was found in 3.0 mg/L or above groups.For 19.5 d uterus,D2 activity decreased and type 3 deiodinase activity increased.The results suggest that excessive iodine has an effect on the embryonic development by regulating maternal-fetal thyroid hormone metabolism.

[Objective]To evaluate the expression of enhancer of zeste homolog 2(EZH2),Ki-67 and intratumoral microvaseu?lar density(iMVD)in human colorectal cancer(CRC),and discuss their relationship with the biological behavior and prognosis.[Methods]The expression of EZH2,Ki-67 and iMVD(labeled by CD34 protein)was measured with immunohistochemical MaxVision method in CRC patients followed up over 5 years.[Results]Positive expression rates of EZH2,Ki-67 and iMVD were 40.91%(45/110),57.27%(63/110)and 46.36%(51/110),respectively. EZH2 expression was positively correlated with distant metastasis(P=0.024). Ki-67 expression was positively correlated with tumor size,infiltration depth,AJCC stage and differentiation(P=0.033~0.015). The iMVD expression was positively correlated with infiltration depth ,AJCC stage and differentiation (P=0.016~0.034). Spearman correlation analysis showed that EZH2 expression was positively correlated with Ki-67 expression(r=0.195,P=0.041), whereas negatively correlated with iMVD expression. Ki-67 expression had a positive correlation with iMVD(r=0.213,P=0.025). Multivariate analysis suggested that the EZH2 expression was an adverse independent factor for survivals of CRC patients(HR 1.965, 95%CI:1.019-3.789,P=0.044 for OS).[Conclusion]EZH2 expression was significantly correlate with the proliferation of tumor cells and may plays an important role in the metastasis and prognosis of CRC.