OBJECTIVE To study the dynamics of nosocomial infections(NI) in our hospital in 2005 and to offer basis for reducing NI by surveillance.METHODS Data of internet report in 2005 were investigated.RESULTS The infection rate of NI in 2005 was 4.27%.The respiratory tract ranked the first place(75.2%);NI occurred mainly in Department of Medicine No.2,cadre ward and infection department;totally 85 isolates of bacteria were found,of which Gram-negative ones accounted for 51.76%.CONCLUSIONS The NI rate is connected with following factors such as age,underlying diseases,days in hospital,and the abuse of antibotics.

ObjectiveTo explore the multidrug resistance(MDR) reversal activity of a novel compound liposome doxorubicin(PLD) and its mechanism.MethodsMTT assay was used to evaluate MDR reversal activity of PLD in P-gp expressing tumor cell lines,KBv200 and MCF-7/ADR.ResultsPLD had a strong reversal in vivo activity,recognized the strong reversal activity than verapamil reversal activity,in 5.0μmol/L concentration of multi-drug resistant cell KBv200 increased sensitivity to vincristine of 45 times.KBv200 PLD-dependent increased in intracellular concentration of rhodamine accumulation(0,2.5,5.0,10μmol/L).Mainly to its cardiac toxicity,bone marrow suppression and hair loss and other side effects were significantly reduced.ConclusionPLD was an efficient modulator,mainly through the continuous accumulation to tumor tissue,tumor local drug concentration,enhanced anti-tumor activity,

0.05).CONCLUSION:Ketoconazole may play its role in treating OHSS by reducing the expression of VEGF.

To investigate the expression change of BDNF in lamina II of spinal cord from partial deafferented cats, L6 segmentsof spinal cord from 20 adult male cats (5 normal cats, 15 unilateral L6 spared roots cats allowed to survive 3 d, 6 d and 12 d re-spectively) were stained with immunohistochemical technique. The results showed: BDNF positive products were mainly dis-tributed in nerve terminals, varicosities and few neurons of spinal cord lamina II in normal cat. After operation, the density ofpositive nerve terminals and varicosities began to decrease on the third day, reached the lowest level on the 6th day and recoveredto normal level on the 12th day on operated side. But the number of BDNF neurons showed no obvious change. The authors sug-gest that the decreased density of BDNF positive products in lamina II on the 3rd and 6th day was related with the degenerationof the nerve fibers and varicosities after section of the adjacent dorsal roots. On the 12th day, the remaining L6 dorsal roots un-derwent collateral sprouting compensatoryly and reestablished functional connection with target neurons. Therefore, BDNF maybe involved in the normal physiological function and the plasticity of spinal cord after damage.

Objective To investigate the effects of extract of Ginkgo biloba leaves EGb761 on 1-methy-l 4-phenylpyridium (MPP+)-induced injury in human neuroblastoma SH-SY5Y cells; To discuss its mechanism of action.Methods Cell culture method was used and SH-SY5Y cell damage model was induced with different concentrations of MPP+ to build Parkinson’s disease model in vitro. The experiment was divided into control group, MPP+ model group, low-, medium-, and high-dose EGb761 groups. The survival rate was determined by MTT assay, and the apoptotic rate was detected by flow cytometry according to AnnexinV apoptosis detection kit. The cell morphology was observed by inverted microscope. NO content in SH-SY5Y cells was detected by Nitric acid reduction method.Results Compared with the control group, the survival rate of SH-SY5Y cells decreased and the apoptotic rate and NO content increased in the model group (P<0.05); Compared with the model group, the survival rate of SH-SY5Y cells increased and the apoptotic rate and NO content decreased in the low-, medium- and high-dose EGb761 groups (P<0.05).Conclusion EGb761 can protect MPP+-induced SH-SY5Y cell from damage by the inhibition of the content of NO free radical.

Objective To observe the effects of low dose irradiation (LDI) on autoiogous CIK cell proliferation, phenotype and killing activity in tumor patients, and to provide the evidence for clinical application of adoptive immunotherapy with CIK cells. Methods Peripheral blood mononuclear cells (PBMC) were separated from 10 patients with malignant tumor, and CIK cells were cultured with different cytokines. (1) After 10 d culture, C1K cells were irradiated with different doses as 30, 50, 80, 100 and 200 Gy of X-rays was also detected. The CIK cell proliferation and killing activity were measured with 3H-TdR incorporation assay and 3H-TdR release assay, respectively and the percentage variation of CD3 +CD56 + were measured with flow cytometry after 24 h. ( 1 ) Autologous CIK cells were irradiated with 80 mGy X-rays. At different culture time ( 12, 24, 48, 72 h) after irradiation, the killing activity was measured with 3H-TdR release assay. (3) The effect of 3d low dose irradiation of 80 mGy X-rays on thekilling activity of CIK cells was also detected. Results After the CIK cells were irradiated with different doses as 50, 80, 100, 200 mGy of X-rays, the CPM values were 20 048.6 ± 2332. 2 ( t = 2.2, P ＜0.05), 21 832.2 ±2975.9 (t=3.5, P＜0.01), 21 000.3 ±2451.1 (t=3.3, P＜0.01), 19908.1 ±2051.0 ( t = 2.2, P ＜ 0.05 ), respectively and the proliferation of CIK cells were significantly higher than that of control group. The CD3 + CD56 + cell percentage of 50, 80, 100 mGy groups were ( 30.3 ±3.8)% (t=2.3, P＜0.05), (32.3±3.4)% (t=4.2, P＜0.01), (29.742.9)% (t = 2.4, P＜0.05 ), respectively. The killing activity of CIK cells of 80, 100 mGy groups were 55.2 ± 5.0 ( t = 3.3, P ＜ 0.01 ), 52.8 ± 4.1 ( t = 2.3, P ＜ 0.05 ), respectively. The killing activity of CIK cells up-regulated significantly at 24 h, dropped to normal levels at 48 h and 72 h. After 80 mGy X-ray irradiation for 3 consecutive days, the killing activity of CIK cells at different time points were 55.2 ± 5.3 (t = 2.6, P ＜0.05),61.9 ± 4.4 (t = 4.7, P ＜0.01), 67.2 ±5.7 (t = 5.7, P ＜0.01) for 24, 48, 72 h,respectively. Conclusions LDI might have the hormesis effect on CIK cells.

AIM: To investigate the anti-cancer effect of cytotoxic T lymphocytes (CTL) activated by sensitized dendritic cells (DCs). METHODS: Immature DCs were induced in vitro from peripheral blood monocytic cells (PBMC) and sensitized by adding tumor cells antigen extract. DCs were identified by their morphology and surface markers. MTT assay was used to evaluate the killing activity of CTL activated by sensitized DCs. The effects of specific CTL cells on inhibiting transplanted tumor HT-29 growth and on preventing HT-29 tumor generation were evaluated by injecting CTL into nude mice. RESULTS: After cultured for seven days, a large number of activated DCs were obtained with typical morphology, extensive stimulatory proliferation capacity and high CD80 (63.5%), CD83 (67.6%) and CD3/HLA-DR (83.2%) expressions. The killing activity of CTL at 20∶1 ratio of effective cells to target cells was more than 75% to tumor cells, 35%-45% to homologous cell line and weaker to other germ cell line (P

Objective To investigate the expression level of thymus microRNA-27a-3p in patients with myasthenia gravis (MG) and to explore the pathogenesis of MG.Methods Thymus tissue samples from 36 cases were collected from December 2014 to February 2015 in the Second Affiliated Hospital of Harbin Medical University.Nineteen thymus tissue samples of MG group were collected from department of chest surgery,17 thymus tissue samples of control group were collected from department of chest surgery or congenital heart disease patients from department of cardiac surgery.The expression of microRNA-27a-3p in the thymus from 36 patients was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR),using U6 as housekeeping control.The Wilcoxon Rank-Sum test was used to analyze the relative expression of microRNA-27a-3p of the two groups.Spearman rank correlation was used to determine the correlation coefficient between microRNA-27a-3p and Quantitative Myasthenia Gravis Score (QMGS).Results (1) The expression level of microRNA-27a-3p in thymus was significantly higher in MG group (0.195(0.049,0.714)) compared with control group (0.045(0.004,0.088),Z =-2.646,P =0.008).(2) Nineteen MG patients were included in the study,out of which 7 were ocular myasthenia gravis (OMG) patients and 12 were generalized myasthenia gravis (GMG) patients.The expression of microRNA-27a-3p in GMG patients (0.493 (0.157,1.123)) was significantly higher than that in OMG patients (0.035 (0.008,0.103),Z =-2.620,P =0.009).(3) There was a positive correlation between the expression of microRNA-27a-3p and QMGS (r =0.576,P =0.010).Conclusions The expression of microRNA-27a-3p in thymus is significantly up-regulated in the patients with MG.MicroRNA-27a-3p may be associated with MG severity and significantly elevated in GMG patients compared with OMG patients.

Objective To investigate the association between myasthenia gravis (MG) and single nucleotide polymorphisms (SNPs) of PTPN22 + 1858C/T,CTLA-4 (+ 49A/G;-1772C/T;-1661A/G),KRAS(rs9226),BCL2(rs4987855) and IGF-1R(rs34804698) genes.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to detect the gene types of SNPs in 76 MG patients who were enrolled in the Second Affiliated Hospital of Harbin Medical University from July 2011 to June 2015 and 59 healthy blood donors.Results In MG patients,the frequences of CTLA-4 +49A/G(rs231775) (57.9％) and-1772C/T (rs733618) (43.4％) were higher than that in the healthy controls (22.1％) (x2 =35.252,P =0.000; x2 =4.098,P =0.043).The frequence of CTLA-4 +49A/G in MG patients combined with thymoma (25.6％) was higher than other subgroups (thymic hyperplasia group:13.8％; normal thymus group:18.4％)(x2 =7.564,P=0.006; x2 =7.155,P=0.007).Meanwhile,the frequence of the C-1772 allele was higher in thymoma group (19.7％) compared with other two groups (thymic hyperplasia group:9.86％ ; normal thymus group:13.8％) (x2 =5.331,P =0.021 ;x2 =4.411,P =0.036).However,the other SNPs were not associated with the risk of MG.Conclusion There are associations of MG with CTLA-4 + 49A/G and-1772C/T SNPs,but not with PTPN,KRAS,BC12 and IGF-1R SNPs.

Objective To construct the prokaryotic recombinant expression vector of PTD4-Cu,Zn-SOD.Methods By using the techniques of gene recombination,the primers of Cu,Zn-SOD and the oligonucleotide sequences of PTD4 were designed,PCR amplification was performed for Cu,Zn-SOD genes,the PCR products were identified,reclaimed and purified,and pET16b served as carrier.The prokaryotic recombinant expression vector of pET16b-Cu,Zn-SOD was constructed using double digestion with Xho Ⅰ and BamH Ⅰ,ligated reaction and plasmid transformation.Then PTD4 gene and pET16b-Cu,Zn-SOD carrier were double digested with Nde Ⅰ and Xho Ⅰ and ligated,and the plasmid was transformed,and the prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD was constructed.The reconstructed vector was analyzed by restriction mapping and was verified by gene sequencing.Results The prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD with a length of 6 207 bp was constructed successfully.The carrier fragment about 5.7 kp and PTD4-Cu,Zn-SOD gene fragment about 510 bp were obtained by double digestion with Nde Ⅰ and BamH Ⅰ,which was consistent with the expected results.The results of gene sequencing showed that the base sequences of pET16b-PTD4-Cu,Zn-SOD were correct when compared with the expected gene sequences.Conclusion The prokaryotic recombinant expression vector of pET16b-PTD4-Cu,Zn-SOD is constructed successfully.