Abstract: Objective　To analyze the phenotype and drug resistance of Robinsoniella peoriensis strains isolated from the blood of patients with prostate cancer and to learn the epidemiological characteristics of the strains. Methods　Culture medium growth characteristics analysis, Gram staining, VITEK MS mass spectrometry identification, in vitro drug susceptibility test, 16S rRNA gene sequencing were performed on the strains, and case summary analysis, historical drug sensitivity results comparison and phylogenetic tree construction were carried out. Results　Four of the repeatability tests of mass spectrometry identification were R. peoriensis, and the identification accuracy was 99.9%, which was the first time that mass spectrometry analysis in China accurately detected this strain. The 16S rRNA gene sequencing confirmed that the strain was R. peoriensis, and GenBank accession number is OL826796. There are currently 18 cases of R. peoriensis related to human infection in the world, mainly including bloodstream infection, prosthetic joint infection, and postoperative wound infection. The homology of OL826796 in this case with HGUE-09/943 (GU322806.1) isolated in Spanish was 99.58%; in vitro drug susceptibility showed that OL826796 was resistant to penicillin and clindamycin, and sensitive to vancomycin, imipenem, tetracycline and metronidazole. Statistical analysis of drug susceptibility of 18 cases found that R. peoriensis could be tested for drug susceptibility by E-test method: penicillin 100% (7/7), clindamycin 70% (7/10), ampenem 0% (0/4), metronidazole 0% (0/9), meropenem 0% (0/4), vancomycin 0% (0/3). Conclusion　R. peoriensis is a rare anaerobic-positive bacillus. When sterile site infection occurs, attention should be paid to timely communication with clinical reports, and penicillin and clindamycin should be used cautiously to fight infection, so as to improve the cure rate of postoperative immunocompromised patients.

Objectives To compare the effect of one-stage revision and two-stage revision for the treatment of culture-negative peripros-thetic joint infection after total hip arthroplasty.Methods A retrospective study was conducted with the clinical data of 41 patients who had chronic periprosthetic joint infection after total hip arthroplasty and then underwent one or two-stage revision surgery from February 2006 to February 2014.The patients were divided into two groups according to different surgical way,namely the 16 patients who received the one-stage revision surgery were regarded as the OSR group,and the other 25 cases who underwent the two-stage revision surgery were regarded as the TSR group.The clinical efficacy of the two surgical way were assessed with Harris Hip score,visual analogue scale (VAS),and rate of infection clearance.Results The average duration of follow up was 29.7 months (9 to 48 months).At the last follow-up,Harris Hip score of TSR group was higher than that of the OSR group,and the difference was statistically significant (P =0.04),and the VAS score of TSR group was lower than that of the OSR group with statistical differences (P =0.02).Additionally,the rate of infection clearance in TSR group was significantly higher than OSR group (P =0.04).Conclusion Culture-negative periprosthetic joint infection can be effectively controled by one or two-stage revision surgery.However,patients got a better prognosis after two-stage revision surgery.

Objective: To observe the migration of 4,4＇-diaminostilbene-2,2＇-disulfonc acid-based fluorescent whitening agents ( DSD-FWAs ) in food packaging paper, so as to provide evidence for quality and safety supervision for paper packaging materials. Methods: Forty-one paper samples with DSD-FWAs positive were made into 6 cm2 pieces and were soaked in four food simulants ( distilled water, 3% acetic acid, 10% ethanol and 95% ethanol, 10 mL each ). The experiment was carried out at the specified soaking temperature and time. The migration amounts of eleven DSD-FWAs were detected by high performance liquid chromatography-fluorescence detection. Results: C.I.220, C.I.24, C.I.210, C.I.85, C.I.113, C.I.264, C.I.353 and C.I.357 were found in all the four food simulants. At the same time and temperature, the migration amount was highest in 10% ethanol, followed by distilled water, 3% acetic acid and 95% ethanol. C.I.220 was dissolved in all four food simulants, in the range of 20-90 ℃, the migration amount increased with soaking temperature; at 20 ℃, 40 ℃ and 60 ℃, the migration amount increased first and then stabilized over time. Conclusion The higher the storage temperature and the longer the storage time of paper packaging, the easier the DSD-FWAs in packaging paper migrate to food.

OBJECTIVETo investigate the impact of 1, 25-(OH)2D3 on left ventricular hypertrophy (LVH) in type 2 diabetic rats.

METHODSType 2 diabetic mellitus (DM) model rats were established by intraperitoneally injecting with 30 mg/kg streptozotocin. After 8 weeks, 19 male rats were identified as diabetic with left ventricular hypertrophy (LVH) by ultrasound examination, and randomly assigned into three groups: untreated (DM-LVH, n=7), treated with insulin (DM-LVH+INS, n=6), and treated with 1, 25-(OH)2D3 (DM-LVH+VD, n=6). Healthy male rats were used as the controls group (n=6). The fasting blood glucose and the insulin level were determined weekly. The left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor level were determined by 4 weeks later.

RESULTSIn the DM-LVH model group, the insulin level was significantly decreased compared with the non-diabetic control group (P<0.05), whereas the blood glucose, left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor expression were significantly increased (all P<0.05). In the DM-LVH+INS and DM-LVH+VD groups, the insulin levels were significantly increased compared with the DM-LVH model group (P<0.05), whereas the other parameters were significantly decreased (all P<0.05).

CONCLUSION1, 25-(OH)2D3 could reverse LVH in diabetic rats and that the mechanism may involve stimulating insulin secretion and reducing blood glucose via direct up-regulation of 1, 25-(OH)2D3-receptor expression.

Animals ; Blood Glucose ; analysis ; Calcitriol ; therapeutic use ; Diabetes Mellitus, Experimental ; blood ; complications ; Diabetes Mellitus, Type 2 ; blood ; complications ; Hypertrophy, Left Ventricular ; prevention & control ; Insulin ; blood ; Male ; Rats ; Rats, Wistar ; Receptors, Calcitriol ; analysis ; Streptozocin

Objective To investigate the impact of 1, 25-(OH)2D3 on left ventricular hypertrophy (LVH) in type 2 diabetic rats.
Methods Type 2 diabetic mellitus (DM) model rats were established by intraperitoneally injecting with 30 mg/kg streptozotocin. After 8 weeks, 19 male rats were identified as diabetic with left ventricular hypertrophy (LVH) by ultrasound examination, and randomly assigned into three groups:untreated (DM-LVH, n=7), treated with insulin (DM-LVH+INS, n=6), and treated with 1, 25-(OH)2D3 (DM-LVH+VD, n=6). Healthy male rats were used as the controls group (n=6). The fasting blood glucose and the insulin level were determined weekly. The left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor level were determined by 4 weeks later.
Results In the DM-LVH model group, the insulin level was significantly decreased compared with the non-diabetic control group (P<0.05), whereas the blood glucose, left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor expression were significantly increased (all P<0.05). In the DM-LVH+INS and DM-LVH+VD groups, the insulin levels were significantly increased compared with the DM-LVH model group (P<0.05), whereas the other parameters were significantly decreased (all P<0.05).
Conclusion 1, 25-(OH)2D3 could reverse LVH in diabetic rats and that the mechanism may involve stimulating insulin secretion and reducing blood glucose via direct up-regulation of 1, 25-(OH)2D3-receptor expression.

Objective To investigate the fear and attitude of the common people to some threatening diseases .Methods Accord-ing to the age and sex structure of the whole nation ,500 common people were randomly enrolled in the public places of Chongqing main urban area .Firstly ,the respondents were asked to name three diseases they feared most .Then their fear degrees to eight im-portant diseases were performed the questionnaire survey .Results The top ten diseases that common people fear most by their own words were cancer ,AIDS ,heart disease ,hypertension ,stroke ,leukemia ,chronic liver diseases ,diabetes mellitus ,dementia and de-pression successively .Meanwhile ,their fear to the eight important diseases ,which was determined by the ratio of people who did fear to all ,were cancer(94 .2% ) ,traffic accidents(89 .2% ) ,heart disease(85 .0% ) ,AIDS(79 .4% ) ,tuberculosis(76 .8% ) ,hepatitis B(76 .6% ) ,Alzheimer′s disease(73 .4% ) and depression(69 .0% ) .Further analysis showed that such fear was significantly influ-enced by age ,sex and education level .Conclusion The knowledge and perception of common people about certain diseases is one-si-ded ,and their health awareness should be improved .

Objective To study the influence of survivin mutant-T34A ( survivinT34A) and survivin deletant-N-terminal 8 amino acids residues ( survivinN-8AA ) on the cell cycle distribution and chemosensitivity in human ovarian cancer HO-8910 cells for explorating the roles of modified survivin-mediated apoptosis induced by chemotherapeutic agents and possible signaling pathways involved. Methods pcDNA3.1 plasmid contained wild-type, survivinT34A and survivinN-8AA genes were transfected into HO-8910 cells,respectively, the control groups were HO-8910 cells transfected with pcDNA3. 1 plasmids. The expression of mRNA was examined by reverse transcription(RT) PCR and identified by DNA sequencing; the cell cycles were determined by flow cytometer analysis ( FCM ); the growth inhibitions rate of cisplatin ( DDP),paclitaxel (PTX) and LY294002 on the transfected cells were determined using methyl thiazolyl tetrazolium (MTT) assay. Results (1) The RT-PCR procedures and genome sequences showed that the survivin mRNA were expressed stable in the transfected HO-8910 cells. (2) There was lower percent of G0/G1 phase cells in SN-HO-8910 cells than that in PC-HO-8910 cells (44. 72% vs. 49.64%, P ＜0. 05) ;while higher percentage of G2/M phase and S phase cells( 1.06% and 54. 22% vs. 0. 56% and 49. 80%, P ＜ 0. 05 ).There was lower the G2/M phase and S phase cells in M-HO-8910 cells 0. 16% and 36. 33%, than that in PC-HO-8910 cells( P ＜ 0. 05 ); while higher percentage of G0/G1 phase cells(63. 51% ,P ＜ 0. 05 ). G0/G1 ,G2/M and S phase cells in Sur-HO-8910 cells were 54. 46%, 0. 62% and 44. 92%, and there were not significantly difference ( P ＞ 0. 05 ), compared to those in PC-HO-8910 cells. ( 3 ) The inhibitory concentration ( IC50 ) of DDP and PTX were higher in Sur-HO-8910 cells than those in control cells [(20. 4 ±6. 1)vs. (14.4 ±3.9)μmol/L,(36.7 ±4.0) vs. (28.6 ±3.6) μmol/L;all P＜0.05]. The IC50 of DDP and LY294002 in SN-HO-8910 cells were lower than those in control cells[(7. 6 ± 1.0) vs. ( 14. 4 ± 3.9)μmol/L, ( 13.2 ± 4. 0) vs. (41.0 ± 7. 9 ) μmol/L; all P ＜ 0. 01]. The IC50 of PTX [( 37. 9 ± 4. 8 ) μmol/L]in SN-HO-8910 cells were higher than that in control cells(P ＜0. 05). The IC50 of DDP in M-HO-8910 cells [(9.9 ± 1.2) μmol/L] were lower than that in control cells(P ＜0. 05) ,and the IC50 of LY294002 in M-HO-8910 cells [(66. 9 ± 4. 8) μ mol/L] higher than that in control cells ( P ＜ 0. 01 ). Conclusions The changes of cells cycle distribution caused by survivinT34A or survivinN-8AA enhanced the G2/M cell cycle-dependent chemosensitivity of PTX. Compared to survivinT34A, survivinN-8AA preferentially to mediate the cytotoxicity of DDP and LY294002, suggesting that it may be related to the cell cycle-dependence of survivin function and to blockage of the formation of its active dimer.

Objective To discuss DNA methylation's effect on pathogenesis of pediatric immune thrombocytopenia (ITP)through detecting the expression level of DNA methyltransferases (DNMTs)mRNA in peripheral blood lymphocytes of children with ITP.Methods Two mL peripheral blood was collected from each of 25 children with persistent and chronic ITP and 20 healthy children (the healthy control group)by using aseptic method in the pediatric ward of the First Affiliated Hospital of Xinxiang Medical University from January 2014 to January 2015.First ethylene diamine tetraacetic acid (EDTA) was used as the anticoagulant.Then separate the mononuclear cells,extract RNA and detect expression levels of DNMT1,DNMT3A and DNMT3B mRNA using reverse transcription-polymerase chain reaction (RT-PCR) method.Results (1) The blood platelet (PLT) of children with persistent and chronic ITP was (36.2 ± 19.6) × 109/L,which was obviously lower than the healthy control group(168.8 ±46.8) × 109/L(t =-11.85,P =0.000).(2)The DNMT1 mRNA expression level of children with persistent and chronic ITP was 0.17 ± 0.05,which was obviously lower than the healthy control group (0.27 ± 0.10) (t =-3.912,P =0.001).The DNMT3A mRNA expression level of children with persistent and chronic ITP was 0.20 ± 0.10,which was obviously lower than the healthy control group (0.32 ±0.11) (t =-3.779,P =0.000).The DNMT3B mRNA expression level of children with persistent and chronic ITP was 0.16 ± 0.1 1,which was obviously lower than the healthy control group (0.31 ±0.11) (t =-4.641,P =0.000).(3) There was positive correlation between the expression of DNMT1 and DNMT3B mRNA(r =0.433,P =0.031).There was positive correlation between the expression of DNMT3A and DNMT3B mRNA(r =0.721,P =0.000).Conclusions (1) Children with persistent and chronic ITP have lower expression levels of DNMT1,DNMT3A,DNMT3 B mRNA,which indicates that DNA methylation contributes to the pathogenesis of pediatric persistent and chronic ITP.(2) DNMTs have synergistic effect on DNA methylation of pediatric persistent and chronic ITP.

Objective To evaluate the sensitivity, repeatability and accuracy of microarray digital PCR system in detecting JAK2 V617F mutation, which was closely related to myeloproliferative neoplasms (MPN).Methods All of the 31 MPN patients with JAK2 V617F mutation, including 18 cases of polycythemia vera(PVs),11 primary thrombocythemias (ETs) and 2 primary myelofibrosis (PMFs), were collected from Huashan Hospital, Fudan University during 2014 -2015, while 10 normal controls and 6 cases with abnormal increased hemoglobin were involved.Human erythroleukemia cell line ( HEL ) and colorectal cancer cell SW480 were used as the mutant and the wild type control, respectively.The sensitivity of microarray digital PCR were verified by detecting the gradient diluted mutation standard harboring 30%, 10%, 1%, 0.1%and 0.01%mutant allele burden, respectively .Repeatability was evaluated by detecting 1%and 10% mutated samples for 5 times, respectively.MGB probe real time PCR was selected as the reference method to verify the accuracy of the digital PCR.Results With digital PCR, the accurate quantitation of JAK2 V617F mutation was achieved down to 0.1%, which is approximate to 0.16 copies per microliter.The results obtained from the two kinds of technique showed a high correlation by linear regression analysis (R2 =0.998 3).The results of repeated samples showed CVs as 17.18% for 1%mutant allele burden and 7.50%for 10%.Among all cases, the 31 patients known mutated were detected as positive and 10 controls as negative by both digital PCR and Real time PCR.In another 6 cases, 2 were found JAK2 V617F mutation of low allele burdens of 0.37% and 0.18% by digital PCR but detected as negative by real time PCR.Conclusions Microarray digital PCR offers a higher sensitivity and better repeatability than real time PCR which could help detect rare JAK2 V617F mutations in MPNs accurately.

OBJECTIVETo observe the changes in Aβ40, Aβ42 and ADDLs in brains of 3 month-old APPswe/PS1dE9 double transgenic mice after six-month intervention with curcumin, in order to discuss the neuroprotective effect of curcumin.

METHODAPPswe/PS1dE9dtg mice were randomly divided into the model group, the Rosiglitazone group (10 mg x kg(-1) x d(-1)) and curcumin high (400 mg x kg9-1) x d(-1)), medium (200 mg x kg(-1) x d(-1)) and low (100 mg x kg(-1) x d(-1)) dosage groups, with C57/BL6J mice of the same age and the same background in the normal control group. After 6 months, the immunohistochemical staining (IHC) and the Western blot method were used to observe the changes in positive cell of Aβ40, Aβ42 and ADDLs in hippocampal CA1 area, their distribution and protein expressions.

RESULTBoth of the immunohistochemical staining and the Western blot method showed more positive cell of Aβ40, Aβ42 and ADDLs in hippocampal CA1 area and higher protein expressions in the model group than the normal group (P < 0.01). IHC showed a lower result in the Rosiglitazone group than the model group (P < 0.05), while Western blot showed a much lower result (P < 0.01). The number of Aβ40, Aβ42 and ADDLs positive cells and the protein expressions decreased in the curcumin high group, the medium group showed a significant decrease (P < 0.01), and the low dose group also showed reductions in the protein expressions of Aβ40 and Aβ42.

CONCLUSIONThe six-month intervention with curcumin can significantly reduce the expressions of hippocampal Aβ40, Aβ42 and ADDLs in brains of APPswe/PS1dE9 double transgenic mice. Whether curcumin can impact Aβ cascade reaction by down-regulating expressions of Aβ40, Aβ42 and ADDLs and show the neuroprotective effect needs further studies.

Alzheimer Disease ; drug therapy ; genetics ; metabolism ; Amyloid beta-Peptides ; genetics ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; Curcumin ; administration & dosage ; Disease Models, Animal ; Hippocampus ; drug effects ; metabolism ; Humans ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Neuroprotective Agents ; administration & dosage ; Plant Extracts ; administration & dosage