Objective To isolate and classigy the bacteriophages specific to Pseudomonas aetuginosa and to investigate biofilm control efficaey of the isolated virulent phages.Methods With P. aeruginosa clinical strains as indicators.bacteriophages were isolated by screening difierent environmental samples.Classification of the isolated phages was done with the methods of restriction fragment analysis of phage genome and host range analysis.Transmission electron microscopy(TEM)was used in phage morphology study.In biogilm control tests,TJC729 was used as the jndicator strain to study the biofilm control efficacy of the isolated phages.Results Total 13 lytic phages specific to P.aeruginosa strains were isolated and named as C1-C13.According to the result of restriction fragment analysis.all 13 phages were double-stranded DNA viruses and could be divided into eight groups.Host range experiments were conducted with 5 laboratory strains and 12 clinical strains of P. aeruginosa.The same infection profiles were observed among phage C1 and C13,C6 and C7,and C9 and C11,respectively.While the remaining 7 phages each had different unique infection profile.Phage C1 was selected randomly to study its morphology.The obtained images showed that phage C1 had an icosahedral head with a non-contractile tail,belonging to the Siphoviridae family.Compared with the single phage,phage cocktail had the best effect on biofilm control.Further experiment results showed that phage C1.C10 and C12 can destroy biofilm after treatment of the biofilm for 24 h.The biofilm amounts were deceased to 32.7%,57.6%and 32.8%of the initial values,respectively.Conclusion Thirteen virulent phages specific to P. aeruginosa had been isolated.The phages could significantly inhibit the biofilm formation and had a certain degree of damage on the biofilm.The results suggested an alternative method for the treatments of P.aeruginosa infections.

AIM:To establish HPLC fingerprint for controlling the quality of Rubus chingii Hu. METHODS: Analysis was performed on Alltech C_(18) column(250 mm?4.6 mm,5 ?m)with a mixture of acetonitrile and water as mobile phase in gradient mode,flow rate was 1.0 mL/min,wavelength at 211 nm. RESULTS: There were 15 common peaks in the HPLC fingerprints of Rubus chingii Hu.The RSD of precision and reproducibility lay within 5%. CONCLUSION: The method has good sensitivity and repeatability.This chromatographic fingerprint method can be used to controll the quality of Rubus chingii Hu.

Objective To observe the serum and cerebrospinal fluid (CSF) levels of procalcitonin (PCT),matrix metalloproteinase-9 (MMP-9) in children with purulent meningitis and viral encephalitis,and evaluat the differential diagnosis value of PCT and MMP-9.Methods The clinical data of 73 patients with intracranial infection admitted to neurology of Qingdao Women and Children's Hospital from September 2014 to December 2015.Twenty-two patients with pumlent meningitis were se1ected as purulent meningitis group,51 patients with viral encephalitis as viral encephalitis group,and another 20 non-infectious convulsion children as a control group.Samples of 2 ml CSF and 3 ml venous blood from all the subjects were collected within 24 hours after admission.The levels of MMP-9 were detected by Enzymelinked immunosorbent assay (ELISA),and the PCT 1evels were measured by electrochemiluminescence immunoassay.The data in multiple groups were compared with analysis of variance,and by SNK-q test to carry on pairwise comparison among groups.The positive rate significance compared by Chi-square test.Linear correlation analysis was used for correlation analysis.Results Mean serum and CSF levels of PCT in the pumlent meningitis patients group were significantly higher than those in the viral encephalitis group as well as the control group (P ＜ 0.01),but the levels of serum or CSF there were no significant difference between the viral encephalitis group and the control group (P ＞ 0.05);Mean serum and CSF levels of MMP-9 in the pumlent meningitis patients group were significantly higher than those in the viral encephalitis group and control group (P ＜ 0.01),and the levels in patients with viral encephalitis were significantly higher than those in control group (P ＜ 0.01);The increased percentage of the serum and CSF PCT in pumlent meningitis group were significantly higher than in viral encephalitis group (P ＜ 0.01).,but there were no significant difference for MMP-9 (P ＞ 0.05);The PCT level of serum or CSF in pumlent meningitis patients was positively correlated with MMP-9 (r =0.498,P ＜ 0.01),but there was no significant correlation in viral encephalitis group (P ＞ 0.05).Conclusion Detection of PCT and MMP-9 level is valuable in differential diagnosis for the pumlent meningitis or viral encephalitis.

Objective To investigate the value of multipoint and multidirectional ultrasound-guided fineneedle aspiration cytology (US-FNAC) in preoperative diagnosis of papillary thyroid microcarcinoma (PTMC).Methods From Jan.2012 to Jan.2013,multipoint and multidirectional US-FNAC were performed in 177 patients with 233 nodules suspected as PTMC,and the cytopathological findings were compared with surgical pathological results and clinical follow-up results.Results Of the 233 biopsies,the cytology were classified as malignant for 146 lesions,suspicious for malignancy for 43 lesions,benign for 24 lesions,follicular lesions for 4 lesions and inadequate sample for 16 lesions.After the second multipoint and multidirectional US-FNAC,only 1 case did not have sufficient cells for diagnosis,and the puncture failure rate was 0.43％.Actually there were 189 malignant and 44 benign lesions confirmed by clinical follow-up and pathological results.The sensitivity,specificity,and accuracy of multipoint and multidirectional US-FNAC was 99.5％,81.8％ and 96.1％ respectively.Conclusions Multipoint and multidirectional US-FNAC has definite value in increasing puncture qualified rate and positive diagnosis rate before operation.

Objective To evaluate the clinical application of multimodality treatment of brain arteriovenous malformation (BAVM) with Onyx embolization and microsurgery.Methods There were 48 patients with BAVM treated by combination of Onyx embolization and microsurgery.The clinical features,therapeutic process,complications and discharge status (by MRI,DSA and Glasgow outcome scale,GOS) were recorded in detail.The prognosis was estimated by modified Rankin scale (mRS) with 6-42 months' follow-up.Results The total resection rate was 97.9％.The therapy-related complications happened in 8 patients (16.7％).Among them,4 cases had dysfunction of central nervous system (8.3％),2 cases with intra-cerebral hemorrhage (4.2％),and 2 suffered from severe cerebral edema (4.2％).Good recovery was seen in 38 cases (79.2％) evaluated by GOS when discharged from hospital.The follow-up result mRS ＜ 3 was 97.9％.Conclusion Multimodality treatment of BAVM with Onyx embolization and microsurgery should be a good therapeutic strategy.

Objective To summarize the clinical and radiological features of DYT6 dystonia with mutations based on the data of our patient cohort as well as the report by others.Methods Clinical data of the 11 patients with DYT6 dystonia in Peking Union Medical College Hospital from June 2009 to May 2012 were retrospectively reviewed and analyzed.Clinical data included gender,onset age,initiative symptom of onset,the sites of involvemet,family history,etc.All patients were examined for brain MRI scan,6 patients were examined for DTI.Results Of the eleven gene-confirmed DYT6 dystonia patients,7 were male and 4 were female,with an onset-age ranged from 5 years to 36 years,the mean age of onset was 19.4years.Eight patients had a family history.There were 10 patients with early onset dystonia and only 1 patient with late onset dystonia.The most common site of onset was the neck (7/11),and the next was the right arm,1-5 body areas were affected at the time of neurological assessment,the average amount was 2.8,and the most frequently affected anatomical site was the neck (10/11),next came lower face,jaw and tongue.Among all the patients,6 patients presented with segmental dystonia,4 patients presented with focal dystonia,only 1 patient presented with generalized dystonia.All the patients with thanatos-associated protein domain-containing apoptosis-associated protein (THAP) domain affected had a family history,but the patients with the same mutant gene varied with clinical manifestation.Only 1 patients with non-THAP domain affected had a family history,but in most families,there were adult asymptomatic mutant gene carriers.Mutations within the THAP domain were associated with an earlier age of onset than non-THAP domain (17.3 and 21.8 years old).Routine MRI of all patients were normal and DTI of 6 patients showed that fractional anisotropy values in the bilateral sensorimotor area in DYT6 dystonia were reduced.A detailed description of a patient with TOR1A and THAP1 gene mutations was given.Conclusions Early onset dystonia is the main manifestation in patients with DYT6 dystonia in China.The most common site of onset is the neck,and the next is the right arm.The most frequently affected anatomical site is the neck,next come lower face,jaw and tongue.Laryngeal dystonia is absent.The patients with same mutant gene show high heterogeneity in the clinical manifestations,mutations within the THAP domain of THAP1 tend to manifest at an earlier age and higher penetration than mutations localized to non-THAP domain.Reduction of fractional anisotropy values indicates that the axonal integrity and coherence in the region of sensorimotor area is damaged in DYT6 dystonia.

Objective To study the profile of peripheral natural killer cells(NK cells)and γδT lymphocytes in human immunodeficiency virus(HIV)infected patients with different disease status and to explore the pathogenesis of acquired immunodeficiency syndrome(AIDS).Methods Three hundred and eleven HIV/AIDS patients without antiviral treatment were enrolled in this study.The percentages and absolute numbers of CD4+T lymphocytes,NK cells,and γδT ceils were measured by flow cytometry.The patients were divided into 3 groups based to their CD4+T lymphocytes counts:low CD4+T lymphocytes group(L),patients with CD4+T lymphocytes <0.20×109L;middle CD4+T lymphocytes group(M),CD4+T lymphocytes counts between 0.20×109and 0.35×109L;high CD4+T lymphocytes group(H),patients with CD4+T lymphocytes counts >0.35×109L.Rank sum test of independent samples of two-group and multiple-group was performed using Mann-Whitney U test and Kruskal-Wallis test.Correlation analysis was done by Spearman and Pearson test. Results The median percentage and cell counts of NK cells(8.4%,103×106L) and γδT cells(3.4%,41×106L)in HIV/AIDS patients were all significantly lower than those of healthy individuals(Z=-5.029,Z=-7.723,Z=-2.437,Z=-6.063;all P<0.01).The median cell counts of CD4+T lymphocytes in L,M,H groups were 0.062×109L,0.276×109L and 0.482×109L,respectively.The median cell counts of NK cells in these 3 groups were 89×106L,97×106L and 146×106L,respectively.NK cell counts were not significantly different between L and M groups,whereas both of them were much lower than that of H group(Z=-3.392,P=-0.001,Z=-4.849,P<0.01,respectively).The median γδT cell counts of L,M and H group were 29×106L,43×106L and 59×106L,respectively.The differences between any 2 groups were not significant.Conclusion These data suggest that the decreasing levels of peripheral NK cells and γδT cells are different after HIV infection.

Objective To investigate the prevalence of primary HIV-1 genotype drug-resistance and viral subtype from 237 treatment-naive patients in China.nethods CD4+ T cell counts,plasma HIV-1 viral load and HIV-1 gene sequencing in total 237 treatment-naive patients enrolled from 20 provinces/regions were detected for the evaluation of primary HIV genotypic drug-resistance.Results The survey of 237 treatment-naive patients from muhicenter areas including Henan Province,Yunan Province,and Shanghai showed that 9 subtypes of HIV-1 strains were finally identified.Most of patients were infected before 2003.Only 3 cases had genotypic mutations associated resistance to antiretroviral drugs,with high resistance to nucleoside reverse transcriptase inhibitors(NRTIs),moderate resistance to protease inhibitors(PIs)and high resistance to non-nucleoside reverse transcriptase inhibitors(NNRTIs).respectively.The prevalence of primary genotypic drug resistance was 1.3%(3/237)in this study.Conclusions The rate of HIV-1 primary genotypic drug-resistance is still relatively low in treatment-naive HIV/AIDS patients while 9 subtypes of HIV-1 strain was diseovered.

To investigate the different reconstitutional profiles for acquired(CD4+ T cell)and innate(NK cell,γδT lymphocyte)immunity after highly active antiretroviral therapy(HAART).Methods The CD4+ CD4+,CD3+ CD4- CD8-,CD3- CD16/CD56+,CD4+ CD45 RA+ CD62 L+ and CD4+ CD45 RA- subsets were measured by flow cytometry.The dynamic changes of these subsets after HAART initiation were assessed in 59 patients who were followed for 12 months in resular 3-month visits.Results At baseline the cell counts of CD4+ T cells including its na(I)ve and memory subsets,NK cell and γδT cells in HIV/AIDS patients were all significantly lower than those of healthy individuals.There was a decrease of 2.33 lg copies/ml in HIV-1 RNA from baseline noted 1 month after initiation of treatment which was sustained through 12 months.CD4+ T cell count showed a bi-phase increase during treatment.The first rapid increase was mainly memory CD4+T cells and this followed by the second slow but steady increase of na(I)ve CD4+ T cells.Increases in NK cell and γδT cell were noted at 3 months of HAART and this restoration were different quantitatively when compared with the oge in CD4+ T cells.Conclusion HAART could induce a different quantitative restorational patterns in peripheral CD4+ T cells,NK cells and γδT cells.

Background To study the pathogenesis and management of diabetic retinopathy (DR) has an important clinical significance.With the development of biomedical photonics in recent years,photobiomodulation therapy has been paid more and more attention.However,the sudy on biological regulation of light to DR is rarely reported.Objective This study was to explore the photobiomodulating effects on the apoptosis of retinal neuronal cells induced by high glucose environment and tried to offer a basis for the management of DR.Methods The retinal neurons were isolated from Wistar rats using immunomagnetic beads and primarily cultured in Neurobasal,and the cells were identified by Nissl staining.The cells were divided into normal control group,high-glucose group and high-glucose+LED group.The glucose at the concentration of 25 mmol/L was added into medium for 48 hours in the high-glucose group,and the cells induced by high-glucose were irradiated in incubator by LED for consecutive 300 seconds per time in a 12-hour interval with the wavelength of 620 nm,maximal power of 1 W,central light radiation exposure of 6.67 mW/cm2 and spot diameter of 2.0 cm.The apoptosis rate of the cells was assayed by flow cytometry;the intracellular Ca2+ content was determined by laser scanning confocal microscope;the relative expression level of phosphorylated serine-threonine kinase (p-AKT) protein in the cells was detected by Western blot.Results The cells grew well 2-3 days after cultured with the polygon and oval shape,and nucleolus were visible.More neuronal processes were obtained in 5-7 days after culture.Nissl staining showed the blue violet color in cytoplasma of neurons.The proportion of neurons and glial cells was 91％.The apoptosis rates of the cells were (7.634±3.176)％,(33.642 ±9.315)％ and (23.914±6.375)％ in the normal control group,high-glucose group and high-glucose+LED group,respectively,and the apoptosis rates of high-glucose group and high-glucose + LED group were significantly higher than that in the normal control group,while the apoptosis rate in the high-glucose+LED group was lower than that in the high-glucose group (all at P＜0.01).The fluorescence of Ca2+ in the cytoplasma was strong in the highglucose group and weak in the normal control group.The fluorescence pixel values in the high-glucose group and highglucose+LED group were significantly higher than that in the normal control group,and that in the high-glucose+LED group was reduced in comparison with high-glucose group (all at P＜0.05).The expressing band of p-AKT protein was strong in the normal control group and weak in the high-glucose group.The relative expressing levels were 10.34± 3.18,2.16±0.46 and 7.15 ±1.72 in the normal control group,high-glucose group and high-glucose+LED group,and relative expression level of high-glucose+LED group was significatly lower than that in the high-glucose group (P＜ 0.05).Conclusions High-glucose environment inhibits PI3K/AKT pathway and calcium homeostasis of retinal neurons,which results in cell apoptosis.Low intensity of LED light irradiation activates the anti-apoptotic PI3K/AKT pathway and therefore reduces apoptosis induced by high glucose.