Objective To analyze the sequence of the novel conjugative plasmid pO157_Sal detected in outbreak isolates of Escherichia coli O157∶H7 .Methods The traE genes of the outbreak isolates in China were amplified by polymerase chain reaction (PCR) and the products were sequenced .The TraE sequences of Escherichia coli O157 ∶ H7 strains from other sources were retrieved from GenBank . Phylogenetic tree based on the TraE sequences was constructed by Neibhor-joining analysis .The whole plasmid sequences of pO157_Sal and pEC4115 were compared .Results The sequences of traE gene were identical among the Chinese isolates . There were homologous sequences of TraE in Escherichia coli O157∶H7 isolates from different sources .Twenty-one out of the 52 pO157_Sal genes were homologous to genes of pEC4115 with amino acid level identity ranging from 28% to 51% .Conclusions Although similar TraE sequences and similar plasmid are found in Escherichia coli O157∶H7 isolates from different sources ,pO157_Sal is only observed in Chinese outbreak isolates .The TraE sequences are conservative among the outbreak isolates ,indicating they are from the same specific source .

Objective To observe the efficacy and safety profile of three-year treatment of HAART in 43 Hemophilia patients co-infected with HIV and HCV. Methods 43 hemophilia patients co-infected HIV and HCV were treated with HAART for 3 years. CD4、 CD3、 CD8 and NK cell counts of the patients were detected by FCM, and the viral load in the plasma was detected with FDA recommended bDNA method. Results The average CD4 count of the 43 patients increased to 257/mm3 (P

Objective To investigate the distribution of human papillomavirus (HPV) type 16 in women cervical infection in eastern Guangzhou, polymorphism of E6/E7 gene and association of gene dosage with disease progression. Methods Flow-through hybridization and gene chips were applied in HPV sub-type identification to screen out HPV-16 positive samples from cervical epithelium samples. HPV-16 E6/E7 gene was amplified through PCR with specific primers. The PCR products were cloned into pMD18-T vectors and fragments were determined through sequencing. Polymorphism analysis were performed through align-ment tools. Fluorescence quantitive PCR were used for the detection of viral E6 gene and L1 gene. Results Thirty-six (4.5%) HPV-16 positive samples were screened out through flow-through hybridization from 806 cervical epithelium samples. HSIL and above happened in 18 (50.0%) of the 36 HPV-16 positive patients. Within E6/E7 gene sequences from 7 selected samples, we found 15 sites with variances and 8 of them would cause coding amino acid change. HIL group (A, 11 cases) and LSIL group (B, 14 cases) possess significantly different gene dosage of both viral E6 gene and LI gene (P <0.05). The ratios of L1/E6 be-tween the 2 groups was not significantly different(P=0.19). Conclusion HPV-16 cervical infection oc-curs in 4.5% women (17-62 years old) in eastern Guangzhou. HIL or above accompany with half of the HPV 16 infected women. Viral load is probably associated with cervical HSIL, though L1/E6 ratios do not suggest viral integration.

Objective To investigate the role of epidermal growth factor receptor and cyclooxygenase-2 pathways in the erlotinib and celecoxib enhanced radiation sensitivity in A549 human lung cancer cell line. Methods IC20 of erlotinib and celecoxibon in A549 human lung cancer cells was measured by MTT assay,Clonogenic assays were used to evaluate the antitumor effects of the drugs and Xirradiation.Flow cytometry was used to assess the apoptosis and cell cycle alteration,and Western blot was used for the detection of Akt and phospho-Akt.Results Both erlotinib and celecoxib could inhibit the proliferation of A549 cells in vitro in a dose-dependent manner and their values of IC20 were (5.15 ± 0.14)and (40.32 ± 1.26) μmol/L,respectively. For radiation survival,the values of Dq,Do,SF2 of the combination of two drugs were lower than those of either drug ( t =6.62,P ＜ 0.05).The SER of celecoxib,erlotinib and their combination were 1.299,1.503 and 2.217,respectively.Flow cytometry assay showed that both celecoxib and erlotinib could enhance radiation-induced G0/G1 arrest,reduce the cell numbmer in S phase,and enhance radiation-induced apoptosis,especially for the combination of drugs.Western blot assay showed that the expressions of Akt protein were similar in all groups.However,pAkt expression was suppresssed by erlotinib and celecoxib,but promoted by radiation.pAkt had the lowest expression in the radiated cells with the treatment of two drugs ( t =4.89,P ＜ 0.05 ).Conclusions The edotinib and/or celecoxib could enhance radiosensitivity probably by increasing cell apoptosis and reducing the number of Sphase cells with low radiosensitivity.

To explore and compare the response of the protease inhibitors or non-nucleoside reverse transcriptase inhibitors-based therapeutic impact on metabolic indices in hepatitis C virus (HCV)/human immunodeficiency virus(HIV) co-infected patients.A randomized,open,and control approach was performed to enroll 273 cases of HCV/HIV co-infected patients on their initial visits and to choose protease inhibitors(PIs group) or non-nucleoside reverse transcriptase inhibitors (NNRTIs group) based therapy treatments for one year.Laboratory results of metabolic indices before and after the treatment were collected.After treatment,the levels of triglyceride in NNRTIs and Pls groups were (1.93 ± 0.99) mmol/L and (1.62 ± 0.93) mmol/L respectively,high density lipoprotein-cholesterol were(1.28 ± 0.55) mmol/L and (1.08 ± 0.53) mmol/L,low density lipoprotein-cholesterol were (2.60 ± 1.44) mmol/L and (2.22 ± 1.16) mmol/L,fasting plasma glucose were (5.92 ± 1.21) mmol/L and (4.79 ± 0.47) mmol/L,serum creatinine were (70.5 ± 14.6) μmol/L and (56.6 ± 8.3) μmol/L,and serum amylase were(66.9 ± 27.5) U/L and(62.7 ± 33.8) U/L respectively.The difference between the two groups was statistically significant(all P＜0.01).There is a therapeutic impact on metabolic indices in patients wtih HCV / HIV co-infection after non-nucleoside reverse transcriptase inhibitors-based regimen.

Objective To establish a model for the calculation of biologically effective dose (BED) and EQD2 (Equivalent dose in 2 Gy fractions) in radioactive seed implantation brachytherapy.Methods The BED formula for EBRT(external beam radiotherapy) and for continuous low dose-rate irradiation established under the L-Q model were introduced.The EDQ2 formula for the continuous low dose-rate irradiation (radioactive seed implantation) was established according to the definition of EQD2 and the formula of BED.The α/β values of common tissues and the Tr 1/2 values reported in the literature were summarized.The EDQ2 formula were further simplified by using the actual values.The empirical formula of EDQ2 for early reaction tissues and late reaction tissues were proposed,named as Wang-Peng empirical formula.EDQ2≈ (10/12) D (Wang-Peng Formula 1) was fit for early response tissue,and EDQ2≈ D/2 (Wang-Peng Formula 2) for late reaction tissues.Further examples on the clinical applications of the proposed formula were given,including primary lung cancer,supraclavicular lymph node metastasis of esophageal cancer and celic lymph node metastasis of cervical carcinoma.Results According to the Wang-Peng empirical formula,the EDQ2 of the late reaction tissue adjacent to the tumor was only about half that of the tumor tissue,so the radioactive seed implantation brachytherapy naturally protected the late reaction tissue by the biological equivalent dose.The actual calculation,showed that the empirical formula of early reaction tissue was more accurate,but the empirical formula of late reaction orgtissue was less inaccurate and could only be roughly estimated.Conclusions The BED calculation formula introduced here and the set of EQD2 calculation formula and Wang-Peng empirical formula established here were theoretically feasible and could be used for the conversion and superposition between the physical dose of radioactive seed implantation brachytherapy and the external irradiation dose.But it should be careful to apply the formula,pay attention to the default conditions,and carefully interpret the calculated results.

Objective To evaluate the efficacy and safety of efavirenz-based therapy in patients with human immunodeficiency virus (HIV)/hepatitis C virus (HCV) co-infection. Methods Fiftythree HIV/HCV co-infected patients received efavirenz-based therapy were followed up for 7 years.The changes of CD4+ T lymphocyte count, HIV virus load, hepatic function, hepatic fibrosis index,blood lipid, blood glucose, blood uric acid and blood routine were observed. The comparison of means before and after treatment was performed by t-test. Results The HIV RNA levels at baseline and endpoint were (4. 56±0. 88) lg copy/mL and (1.70±1.10) lg copy/mL, respectively (t=14. 781, P＜0.01). The peripheral blood CD4+ T lymphocyte counts were ( 188.37±151.14)×106/L and (445.18±314.25)×106/L, respectively (t=5.362, P＜0.01).The alanine aminotransferase (ALT) levels were (36.6±16.3) U/L and (57.2±9.9) U/L, respectively (t=7.864, P＜0. 01).The glycocholic acid levels were (444.22±476.74) mg/L and (556.88±733.05) mg/L, respectively (t=0.938, P＜0.05). The Ⅳ-collagen(Ⅳ-C) levels were (45.13±8.25) ng/mL and (47.88±4.51) ng/mL, respectively (t= 2.129, P＜0.05). The riacylglycerol levels were (1.57±0.65)mmol/L and (2.51±1.29) mmol/L, respectively (t=4.737, P＜0.01). The blood uric acid levels were (298.5±48.2) mmol/L and (495.1±89.4) mmol/L, respectively (t= 14.092, P＜0.01).Conclusions The efavirenz-based therapy is efficacious in HIV/HCV co-infected patients, but it could cause liver injury and metabolic disorder.

To demonstrate the fructose metabolism pathway in Bifidobacterium Longum NCC2705 and to construct its fermentation model, we explored the comparative proteome cultivating the strain on glucose or fructose, based on a proteomic reference map of B. longum NCC2705 constructed earlier. Then, we used matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry and electro-spray ionization tandem mass spectrometry (ESI-MS/MS) for differently expressed proteins identification. Furthermore, with semi-quantitative RT-PCR we determined the distinctively expressed proteins at the level of transcription. Proteomic comparison of glucose- and fructose-grown cells demonstrated much similarity. On the page of fructose there were all the enzymes and proteins that exist during the process of glucose degradation. We observed a greater variation of more than three-fold for the identified 9 spots representing 5 protein entries by MALDI-TOF MS. The sugar-binding protein specific to fructose (BL0033) and an ABC transporter ATP binding protein (BL0034) showed higher expression level from cells grown on fructose. It was also determined by semi-quantitative RT-PCR subsequently. BL0033 time course and concentration experiments showed that the induction time correlated to higher fructose concentration, and increased expression of BL0033. Fructose was catabolized via the same degradation pathway as glucose at the level of proteomics. BL0033 was induced by fructose. All results suggest that the uptake of fructose into the cell may be conducted by a specific ABC transport system, in which BL0033 and BL0034 as components might have played an important role.
Bifidobacterium ; chemistry ; genetics ; metabolism ; Culture Media ; Fermentation ; Fructose ; pharmacology ; Glucose ; pharmacology ; Proteome ; analysis ; genetics ; Proteomics ; methods

Objective To study the changes of serum brain natriuretic peptide(BNP),uric acid(UA)and procalcitonin(PCT)levels in children patients with dilated cardiomyopathy(DCM).Methods 24 children pa-tients with DCM and 24 children undergoing healthy physical examination from March 2015 to September 2016 were selected as the observation group and control group.The observation group was divided into differ-ent Ross grade groups according to the improved Ross heart failure grade scoring.The levels of serum BNP, UA and PCT in each group were detected and their correlation with the main echocardiographic indicators was analyzed.Results The levels of BNP,UA and PCT had statistical difference between the observation group and control group(P<0.05);the BNP and UA levels had statistical difference among different Ross grades (P<0.05),moreover which were increased along with the severity increase,but the PCT level had no statisti-cally significant(P> 0.05);the BNP level was positively correlated with the DCM severity(r= 0.713,P=0.000),the UA level was positively correlated with the DCM severity(r=0.489,P=0.002),while the corre-lation between the PCT level and DCM severity had no statistical significance(r=0.288,P=0.076);the BNP had obvious correlation with main echocardiographic indicators(P<0.05).Conclusion The levels of serum BNP,UA and PCT in children patients with DCM are changed significantly,in which BNP and UA may be the good markers for reflecting the disease severity and progression or prognosis.

Objective To investigate the utilization rate of gold fiducial markers and reasons for abandonment of gold fiducial markers in the CyberKnife VSI System, and to provide reference data for implantation of gold fiducial markers and radiotherapy planning. Methods From March to August,2017,a total of 47 patients had gold fiducial markers implanted or pasted. In those patients, 42 patients had gold fiducial markers implanted,including 32 receiving computed tomography(CT)-guided 3D-printing coplanar template assisted implantation, 1 receiving CT-guided 3D-printing non-coplanar template assisted implantation,1 receiving CT-guided implantation,and 8 receiving ultrasound-guided implantation. A total of 44 patients received the CyberKnife treatment, including 2 patients who failed to use gold fiducial markers and were treated with spine tracking instead and 3 patients missing the treatment for other reasons. The numbers of utilized and abandoned gold fiducial markers were recorded for calculation of the utilization and abandonment rates. The reasons for abandonment of gold fiducial markers were analyzed and classified. Results A total of 134 gold fiducial markers were implanted into or pasted to the 44 patients.In all the gold fiducial markers, 111 were utilized and 23 abandoned, yielding a utilization rate of 82.8% and an abandonment rate of 17.2%.The reasons for abandonment of gold fiducial markers included large rigidity error(26.1%), unqualified implanted fold fiducial markers(17.4%), displacement of gold fiducial markers(26.1%), and others(30.4%). Conclusions Compared with the CT-guided or ultrasound-guided implantation of gold fiducial markers, the CT-guided 3D-printing coplanar or non-coplanar template assisted implantation of gold fiducial markers requires only two puncture needles for each implantation and implants two gold fiducial markers by a single needle,which reduces the number of puncture needles,risk of puncture-induced injury,and incidence of complications after implantation. Not all the gold fiducial markers implanted by a variety of ways will be utilized. Some gold fiducial markers will be abandoned for different reasons,which should be taken into account during implantation of gold fiducial markers.