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The common clinical subtypes of juvenile idiopathic arthritis (JIA) include systemic onset juvenile idiopathic arthritis (SOJIA), oligoarthritis/polyarthritis juvenile idiopathic arthritis and juvenile spondyloarthritis. Juvenile idiopathic arthritis has no specific diagnostic index, and needs to be differentiated from infectious diseases and malignant diseases. The onset of SOJIA is rapid, the disease progresses rapidly, and it is easy to be complicated with macrophage activation syndrome (MAS) which is life-threatening. The experience of pediatric rheumatologists in dealing with JIA is still insufficient, and the standardized diagnosis and treatment level of this disease needs to be further improved. Based on the experience and guidelines of diagnosis and treatment in China and abroad, we formulated this diagnosis and treatment standard, aiming at standardizing the diagnosis and treatment of the subtypes of JIA and MAS, so as to reduce the incidence of disability and serious complications and improve the prognosis.
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Objective To observe the long-term efficacy and safety of autologous hematopoietic stem cell transplantation (AHSCT) for systemic sclerosis (SSc) patients.Methods Between May 2007 and June 2009,4 patients with SSc were enrolled in the study.Peripheral blood stem cells were mobilized with cyclophosphamide (CTX) followed by granulocyte colony stimulating factor (G-CSF).Conditioning was performed with i.v.cyclophosphamide 50 mg ·kg-1 ·d-1 for 4 days.The results of the modified Rodnan skin score (mRSS),thoracic high-resolution computer tomography and pulmonary function were collected after transplantation.Results There was an improvement in mRSS,lung function and HRCT in the six months after AHSCT.Within six month to one year after transplantation,one patient had sustained and two patients recurred.After active treatments two patients were improved again.During the follow-up of 8.7 (4.1-9.8) years,three patients were stable and one patient died.Infection and hepatic function injury were the major complications.There was not transplant-related mortality.Conclusion AHSCT with CTX as a pre-conditioning regimen is safe and effective for SSc.The efficacy for patients with short course,rapid progress and edema is significant.However,long-term efficacy is poor,and long-term maintenance treatment is needed.
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Objective@#To observe the long-term efficacy and safety of autologous hematopoietic stem cell transplantation (AHSCT) for systemic sclerosis (SSc) patients.@*Methods@#Between May 2007 and June 2009,4 patients with SSc were enrolled in the study. Peripheral blood stem cells were mobilized with cyclopho-sphamide (CTX) followed by granulocyte colony stimulating factor (G-CSF). Conditioning was performed with i.v. cyclophosphamide 50 mg·kg-1·d-1 for 4 days. The results of the modified Rodnan skin score (mRSS), thoracic high-resolution computer tomography and pulmonary function were collected after transplantation.@*Results@#There was an improvement in mRSS, lung function and HRCT in the six months after AHSCT. Within six month to one year after transplantation, one patient had sustained and two patients recurred. After active treatments two patients were improved again. During the follow-up of 8.7 (4.1-9.8) years, three patients were stable and one patient died. Infection and hepatic function injury were the major complications. There was not transplant-related mortality.@*Conclusion@#AHSCT with CTX as a pre-conditioning regimen is safe and effective for SSc. The efficacy for patients with short course, rapid progress and edema is significant. However, long-term efficacy is poor, and long-term maintenance treatment is needed.
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Objective To compare the effectiveness and safety of endoscopic submucosal dissection ( ESD) with endoscopic piecemeal mucosal resection ( EPMR) for early esophageal cancer and precancerous lesions with length more than 5 cm. Methods A retrospective analysis was performed on data of 85 patients diagnosed as early esophageal cancer and precancerous lesions with length more than 5 cm in Fujian Medical Association of Early Esophageal Carcinoma from January 2012 to July 2017. The patients were divided into ESD group (52 cases) and EPMR group (33 cases), and the effectiveness and safety between the two groups were compared. Results There was no significant difference on the complete resection rate between the two groups[86. 5％ (45/52) VS 87. 9％ (29/33), P>0. 05]. The operative time (58. 53±30. 50 min VS 32. 06±9. 12 min), postoperative fasting time (4. 18±1. 30 d VS 3. 67±0. 96 d), postoperative hospital-stay time (7. 45±2. 44 d VS 6. 54±1. 73 d), and postoperative antibiotics using time (3. 48±2. 33 d VS 1. 96±2. 20 d) in ESD group were higher than those in EPMR group (all P<0. 05). There were no significant difference in the rate of intraoperative complication and short-term postoperative complication, such as fever, chest pain, and postoperative bleeding, between the two groups ( all P>0. 05 ) . But the postoperative stricture rate of ESD group was higher than that of EPMR group[23. 1％ (12/52) VS 6. 1％(2/33), P<0. 05]. During the follow-up of 3-63 months, 5 cases recurred in ESD group and 1 case in EPMR group, with no significant difference ( P>0. 05). Conclusion ESD and EPMR have equivalent efficacy and safety on the treatment of early esophageal cancer and precancerous lesion. EPMR has a shorter operative time, lower rate of post-operative stricture, and is easier to master.
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Objective To observe the efficacy and safety of rituximab in the treatment of systemic sclerosis (SSc).Methods This was a prospective,non-randomized controlled trial.All patients with SSc were hospitalized from January 2011 to August 2015.Fifty-two patients were enrolled,including 15 patients in the intervention group and 37 patients in the control group.Both groups were given cyclophos-phamide and prednisone.The intervention group was also given rituximab 375 mg/m2,once every 4 weeks,a total of 4 treatments.The erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),intedeukin 6 (IL-6),modified Rodnan skin score (mRSS),HRCT,forced vital capacity (FVC％),1％ forced expiratory volume percentage (FEV 1％),percentage of carbon monoxide dispersion percentage (DLCO％) were assessed alternatively before treatment,6 months and 12 months after treatment.The repeated measures analysis of variance and t-test were used for statistical analysis.Results Both groups showed improvement in ESR,CRP,IL-6,mRSS,HRCT,FVC％,FEV1％ and DLCO％ (P＜0.05),but ESR [6 month after treatment (33±9) mm/1 h,12 month after treatment (20±4) mm/1 h],CRP [6 month after treatment (12.7±3.4) mg/L,12 months after treatment (12.7± 3.4) mg/L],IL-6 [6 month after treatment (73±10) pg/ml,12 month after treatment (57±11) pg/ml],mRSS [6 months after treatment (22.2±1.3),12 month after treatment (18.4±3.1)],HRCT score [6 month after treatment (11.9±1.4),12 month after treatment (11.3±1.0)],in patients of the intervention group were decreased more significantly than patients of the control group (P＜0.05),and FVC％ [6 month after treatment (69.0± 3.4)％,12 month after treatment (77.2±4.1)％],FEV1％[6 month after treatment (73.3±3.4)％,12 month after treatment (78.4±1.6)％] and DLCO％ [6 month after treatment (60.6±2.7)％,12 month after treatment (70.7± 3.0)％] were increased more significantly than patients of the control group (P＜0.05).It showed that the efficacy of the intervention group was better than the control group.There was no infusion reactions,infection,hepatitis B virus reactivation and other side effects in the treatment were observed.Conclusion Rituximab can reduce the level of inflammation in patients with SSc and improve skin sclerosis and lung function.Rituximab combined cyclophosphamide is expected to be a new,safe and effective treatment of SSc.
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Objective To investigate the transcriptional autoregulation of PhoP/PhoQ under different growth conditions in Yersinia pestis.Methods The entire promoter region of YPO1635 was amplified and cloned into the pRW50 vector containing a promoterless lacZ reporter gene.The recombinant LacZ reporter plasmid was transformed into the wild-type strain (WT) and the phoP mutant strain (ΔphoP),respectively,to measure the promoter activity (the β-galactosidase activity) of the target gene in WT and ΔphoP by using the β-galactosidase enzyme assay system.Total RNAs were extracted from WT and ΔphoP strains,and primer extension assay was employed to detect the promoter activity by examining the amount of primer extension products of YPO1635 in WT and ΔphoP.Results The LacZ fusion results showed that the transcription of YPO1635 was positively regulated by PhoP under L-TMH and brain-heart infusion(BHI) conditions,but it was not regulated in H-TMH medium.The primer extension assay detected two transcriptional start sites located at 90 and 118 bp upstream of the translation initiation site of phoP,named P1 and P2,respectively.Under low Mg2+ TMH conditions,the promoter activity of P1 rather than P2 was positively regulated by PhoP.Under high Mg2+ TMH conditions,the promoter activities of both P1 and P2 showed no obvious difference in the WT and ΔphoP strains.Under rich BHI conditions,both promoters were under negative control of PhoP.Conclusion Different autoregualtion patterns of PhoP/PhoQ under different growth conditions would help Y.pestis to quickly adapt to the changing living environment.
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Objective To investigate the influence of early growth response gene-1 (EGR1) on the autophagy of host cells following infection with human T cell leukemia virus type 1 (HTLV-1).MethodsA HTLV-1-positive cell line MT2 was co-cultured with HeLa cells for 24 h to construct the virus early infection model.Immunoblotting assay was used to detect the expression of HTLV-1 core protein p19 and EGR1.Luciferase reporter gene analysis was used to detect the transcriptional activity of 5′-regulatory sequence of EGR1 at different time points after co-culturing.An effective small interfering RNA (siRNA) targeting EGR1 was screened out and transfected into HeLa cells by Lipofectamine 2000.Then the transfected HeLa cells were co-cultured with the HTLV-1-positive cell line MT2 for 24 h.Immunoblotting assay was used to detect HTLV-1 core protein p19, EGR1 and autophagy-related protein LC3.Real-time PCR was performed to detect viral load.Autophagosome was analyzed by immunofluorescence after co-culturing.Results The expression of EGR1 and the transcriptional activity of pEGR1-luc gradually increased after co-culturing HeLa cells with MT2 cells for 8 h (P<0.01).The expression of EGR1 was positively correlated with host cell autophagy following HTLV-1 infection.The effective siRNA for silencing the expression of EGR1 was obtained and named as siE2.The viral load, the expression of HTLV-1 core protein p19 and the proportion of LC3B/LC3A in the co-culture model were markedly down-regulated by RNA interference with siE2, which was concomitant with a persistent decrease of intracellular autophagosome (P<0.01).Conclusion EGR1 is associated with host cell autophagy and viral replication in HTLV-1 infection.
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Objective To study the transcriptional regulation of vp1667 by H-NS in Vibrio parahaemolyticus.Methods Total RNAs were extracted from Δhns and WT strains.Quantitative RT-PCR was carried out to calculate the transcriptional variation of vp1667 between Δhns and WT.Primer extension assay was also employed to detect the transcription start site and the promoter activity (i.e.the amount of primer extension products) of vp1667 in Δhns and that in WT.The promoter DNA region of vp1667 was amplified, purified, and cloned into the corresponding restriction endonuclease sites of pHRP309 that harbors a gentamicin resistance gene and a promoterless lacZ reporter gene.The recombinant pHRP309 plasmid was transformed into Δhns and WT, respectively, while β-galactosidase activity in cellular extracts was measured using a β-galactosidase enzyme assay system.The over-expressed His-H-NS was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns.The electrophoretic mobility shift assay (EMSA) and DNaseⅠ footprinting were then applied to analyze the DNA-binding activity of His-H-NS to vp1667 promoter region in vitro.Results and Conclusion The primer extension assay detected one transcription start site for vp1667, which was located at 28 bp upstream of vp1667, and its transcribed activity was under the negative control of the H-NS.The EMSA and DNaseⅠ footprinting assay results showed that His-H-NS was unable to bind to the promoter-proximal DNA region of vp1667, suggesting that H-NS indirectly inhibits the transcription of vp1667.
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Objective To study the impact of QseBC on the motility of Salmonella enterica serovar Typhi ( S.Typhi ) . Methods The motility of wild-type ( WT) and null mutants (ΔqseB and ΔqseC) at mid-log phase was investigated by swimming assay.Quantitative RT-PCR was carried out to calculate the transcriptional variation of flhD and qseB among WT,ΔqseB andΔqseC.QseB overexpressing strain was constructed to compare its motility and flhD expression with the wild-type control.Results The result of motility assay showed that the motility of ΔqseB was similar to that of the WT strain , while the motility of ΔqseC was much lower than that of WT .qRT-PCR revealed that compared with WT , the expression of flhD was significantly decreased in ΔqseC while the expression of qseB was increased considerably .The motility of QseB overex-pressing strain was lower .Conclusion The expression of flhD may be regulated by QseBC which has an effect on the motil-ity of S.typhi, and the overexpression of QseB may inhibit the motility .
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Objective To investigate the clinical characteristics of Beh(c)et's disease (BD) complicated with cerebral venous sinus thrombosis (CVST).Methods In patients data at Peking Union Medical College Hospital between February 2004 and April 2013 were reviewed retrospectively to identify patients who were diagnosed as BD complicated with CVST.SPSS 19.0 software was used for statisticals analysis.Data of normal distribution was expressed by x±s (standard deviation) and that of abnormal distribution by median and range.Results Of 601 inpatients with BD during that period,12 patients (2％,6 male and 6 female) developed CVST,and their age was (28±10) years old (range 15 to 52 years old).Most of patients (92％,11/12)suffered neurological symptoms after other initial systemic symptoms of BD in mean (56±35) months.The most frequent neurological signs were headache (11 patients,92％),papilledema (9 patients,75％),nausea/vomiting (8 patients,67％) and blurred vision (6 patients,50％),with progression as the main mode of onset (9/11,82％).Eight out of 10 patients (80％) were found elevated cerebrospinal fluid (CSF) pressure by lumbar puncture.Occlusions of multiple sinuses (8 patients,67％) were more frequent than single occlusion (4 patients,33％).The most frequent locations of CVST were the transverse sinus (9 patients,75％),superior sagital sinus (8 patients,67％) and sigmoid sinus (7 patients,58％).Other than CVST,8 patients were complicated with other sites of thrombosis (7 cases in lower extremity veins and 1 in pulmonary artery).In addition,cerebral infarction was found in 3 patients,subarachnoid hemorrhage and right temporal lobe hemorrhage in 1 patient,respectively.After glucocorticoid,immunosuppressant,as well as anticoagulant and dehydration therapy,with a median follow-up of 8 months (0.7 to 88.8 months),10 patients achieved clinical improvements (83％),2 patients relapsed (17％).Conclusion CVST is a relatively rare complication of Beh(c)et's disease,with headache,papilledema,nausea/vomiting and blurred vision as the common clinical manifestations.By early identification,timely diagnosis and active treatment,the prognosis of most patients is good.
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Objective To investigate the quinolone resistance determinants in ciprofloxacin-resistant Acinetobacter bau-mannii (ABA)clinical isolates.Methods One hundred and fourteen ciprofloxacin-resistant ABA strains were collected from six Chinese hospitals .The quinolone resistance determining region ( QRDR) of 4 target genes ( gyrA, gyrB, parC and parE) was amplified , sequenced and compared with the reference genome of ATCC 17978 to identify possible resistance-related mutations.Nine plasmid-mediated quinolone resistance (PMQR) genes (qnrA, qnrB, qnrC, qnrD, qnrS, qepA, aac(6′)-Ⅰb-cr, oqxA and oqxB) were also amplified, and the amplicons were then sequenced to determine their character-istics.Results Almost all isolates (113/114, 99.1%) harbored a substitution in codon 83 of gyrA gene, leading to a Ser83Leu mutation.Meanwhile,58.8%(67/114) of the isolates possessed dual mutations of GyrA-Ser83Leu and GyrA-Ser80Leu, which were known determinants for ciprofloxacin resistance .There were also multiple non-synonymous substitu-tions in gyrB, leading to Arg393Ser, Arg393Cys, Thr401Ala, Pro406Ser, Val430Phe, Cys440Ser and Gly480Arg muta-tions with prevalence rates of 95.6%, 0.9%, 96.5%, 96.5%, 100%, 96.5%and 96.5%,respectively.For parE, all the seven mutations were synonymous and found in more than 96%of the tested isolates.For PMQR genes, although 83.3%(95/114) of the isolates were positive for aac(6′)-Ⅰb, nocrmutations were identified.None of the other eight PMDR genes were found in our strain collection .Conclusion Although multiple mutations are identified in gyrB and parE, these mutations might be the characteristic SNP markers for specific clones , unlikely linked to quinolone resistance .No PMQR is found in the tested isolates.Mutations in chromosomal QRDR (GyrA-Ser83Leu and ParC-Ser80Leu) are the main determi-nants of ciprofloxacin resistance in our ABA collection .
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In order to investigate the effect of T-bet on malignant cells, we selected SGC-7901, a kind of human gastric carcinoma cell line, and used gene clone technique and adeno-associated virus (AAV) packing technology, thus obtaining a recombinant rAAV-eGFP-T-bet and T-bet gene-transfected SGC-7901 cells. Then the function of T-bet gene-infected SGC-7901 cells was researched by detecting the levels of IFN-gamma and T-bet production. The results showed: (1) It was verified that rAAV-T-bet's packing was completed; (2) After SGC-7901 cells was transfected by rAAV-eGFP-T-bet, a green fluorescence was found in about 30%-40% SGC-7901s, and the gene of 1670 bp (T-bet) and 388 bp (IFN-gamma) were generated from SGC-7901s cells; (3) The proteins of IFN-gamma and T-bet secreted by SGC-7901 cells were also detected. These reveal that SGC-7901 cell is efficiently infected by rAAV encoding T-bet, which can induce transfected cells to secret IFN-gamma. It may be useful in the researches on cancer immune therapy of transfecting T-bet gene.
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Humans , Cell Line, Tumor , Dependovirus , Genetics , Metabolism , Green Fluorescent Proteins , Interferon-gamma , Recombinant Proteins , Genetics , Stomach Neoplasms , Genetics , Metabolism , T-Box Domain Proteins , Genetics , TransfectionABSTRACT
Objective To investigate the antimicrobial susceptibility of Pseudomonas aeruginosa (P. aeruginosa) isolated from Zhenjiang area to 13 routinely used antibiotics and identify the structure and dissemination of class Ⅰ integron. Methods K-B test was used to determine the resistant rate of 71 strains of P. aeruginosa. DNA template was extracted by boiling method, PCR method was utilized to detect class Ⅰintegron, and subsequently gene cassettes were analyzed by sequencing. Results The resistant rates to 13 routinely used antibiotics were quite different from 18. 3 to 77.5% among 71 strains of P. aeruginosa. The prevalence of class Ⅰ integron was 38%. These integrons include 5 gene cassettes ( aadB, aac (6) - Ⅱ , PSE-Ⅰ , dfrA17 and aadAS), in which dfrA17 and aadA5 gene cassette were frequently found. Comparing with the negative strains of integron, the positive strains of integron has obviously higher resistance to ten the antibiotics including piporacillin, piperacillin-tazobactam, ceftriaxone, cefepime, ceftazidime, gentamicin,amikacin, tobmmycin, levofloxacin, and ciprofloxacin. Conclusions The resistant rates of P. aeruginosa to 13 drugs were different, and the resistant rates of integron positive strains were obviously higher than integron negative strains, which indicates that integron may play an important role in multidrug reisistance of P. aeruginoosa.