Objective To analyze the clinical manifestations of refractory mycoplasma pneumoniae pneumonia (RMPP)which unresponded to methylprednisolone in the dosage of 2 mg·kg-1 ·d-1 for 3 day.Methods Retrospective analysis was performed on the clinical data of 120 children with RMPP.The patients were divided into effective group and ineffective group according to ini-tial effeet of 2 mg· kg-1 · d-1 methylprednisolone.The elinical manifestations,laboratory examination,radiological features and bronchofibroscopic findings of the children were compared.Results Twenty-eight patients in 120 were poor curative effect after regular 2 mg·kg-1 ·d-1 methyl prednisolone therapy,accounted for 23.3%.There were 10 patients in 28 with mixed infection,ac-counted for 35.7%;13 patients with appeared necrotizing pneumonia,accounted for 46.4%;13 patients with secretion obstruction, accounted for 46.4%;10 patients with endometrium necrosis,accounted for 35.7%.92 patients in 120 were good curative effect af-ter regular 2 mg·kg-1 ·d-1 methyl prednisolone therapy,accounted for 76.7%.There were 5 patients in 92 with mixed infection, accounted for 5.4%;10 patients with secretion obstruction,accounted for 10.9;5 patients with secretion obstruction,accounted for 5.4%.The difference was statistically significant (P <0.05).There were correlation between effect of hormone therapy and the levels of WBC,N,hs-CRP,LDH,PCT,IL-6,IL-8,LP,SF,D-dimmer.Multiple factors analysis showed that N,hs-CRP,LDH,IL-8, IL-6 were independent risk factors influence the effect of regular hormone therapy (P <0.05).The effective were improved after in-creasing hormone doses or share ivig.Antibiotic use days in effective group was obviously lower than that in ineffective group,.The difference was statistically significant (P <0.05).Duplex control antibiotics use ratio in ineffective group was significantly higher than that in effective group.The difference was statistically significant (P <0.05).Conclusion Treatment with 2 mg·kg-1 ·d-1 methvlprednisolone could improve clinical symptoms and radiological manifestations of most children with RMPP quickly.But it may be ineffective in some situations such as N,hs-CRP,LDH,IL-8 and IL-6.

In recent years, with the advances in life cycle of hepatitis C virus (HCV) and the approval of direct-acting antiviral agents (DAAs), great progress has been made in treatment of chronic hepatitis C (CHC).This article reviews the research progress of DAAs in CHC management, showing the possibility of the elimination of HCV infection by DAAs.DAAs have not been widely clinically used due to high medication costs, but can be used as a supplement for the combination therapy of pegylated interferon and ribavirin.

Objective To investigate the signaling mechanisms underlying synergistic induction of MUC5AC mucin by nontypeable Haemophilus influenzae(NTHi)and epidermal growth factor (EGF).Methods The expression of MUC5AC was measured by real-time quantitative polymerase chain reaction(PCR)and Luciferase assay.Western blot was performed to examine the synergistic induction of phosphorylation of P38,extracellular signal-regulated kinase(ERK)and P21-activated kinase(PAK)4 or the effect of dominant negative mutant of PAK4 on the synerglstic induction of phosphorvlation of P38 mitogen-activated protein kinase(P38MAPK)and ERK in HM3 cells treated with NTHi and EGF.Luciferase asgay was also performed to examine the effect of P38,ERK inhibitors or dominant negative mutants of P38MAPK and ERK on synergistic enhancement of NTHi-induced MUC5AC up-regulation by EGF at transcriptional level.Real-time quantitative PCR was performed to examine the effect of PAK4 siRNA on synergistic induction of NTHi-induced MUC5AC up-regulation by EGF.ResuIts NTHi induced MUC5AC mucin expression at both mRNA and transcriptional levels.Synergistic induction of phosFIhorylation of P38MAPK,ERK and PAK4 were observed in HM3 cells treated with NTHi and EGF.Either SB203580,a specific inhibitor for P38MAPK or PD98059,a specific inhibitor for ERK inhibited synergistic induction of MUC5AC at transcription level.Furthmore, overexpressing dominant negative mutant of P38MAPK and ERK also inhibited synergistic induction of MUC5AC at transcription level. PAK4 siRNA inhibited the synergistic induction of MUC5AC by NTHi and EGF. Overexpressing dominant negative mutant of PAK4 also reduccd synergistic induction of phosphorylation of P38 and ERK. Conclusion Synergistic induction of MUC5AC mucin by NTHi is up-regulated by EGF via PAK4-dependent P38MAPK and ERK pathways.

There are about 150 million people around the world with chronic hepatitis C virus (HCV)infection currently,of whom 20%will ultimately progress to cirrhosis and eventually die of end-stage liver disease and hepatocellular carcinoma.Interferon (IFN)has long been recognized as the cornerstone of the treatment of chronic hepatitis C because of its role in sustained virologic response and prevention of disease progression.However,it has limited efficacy and multiple adverse effects.In recent years,direct-acting antiviral agents (DAAs) have shown good efficacy.This review summarizes the recent advances in IFN-free anti-HCV therapeutic regimens based on DAAs.We believe that,with the emergence of DAAs,IFN-free therapies will develop rapidly and display better prospects.

Objective To investigate the different biological characteristics of high-and low-expression of ATP-binding cassette transporter(ABCG2)in human multi-drug resistant nasopharyngeal carcinoma CNE2/DDP cell line(abbreviated as ABCG2High cell and ABCG2Low cell),and to analyze drug-resistant characteristics of the two cell lines.Methods ABCG2High and ABCG2Low cells were isolated by flow cytometry,while their cloning efficiency,cell cycle distribution,and the expression of ABCG2 before and after the 5th passage were examined as well.mRNA expressions of drug-resistant genes,such as ABCG2,Bcl-2,MDR1,MRP and MGMT,were detected by reversed transcriptive polymerase chain reaction(RT-PCR).MTT assay was utilized to detect the light absorbance(A)from day 1 to 14 to determine growth kinetics.Moreover,the drug sensitivity of the two cell lines to fluorouracil,cisplatin,vincristine,carboplatin,epirubicin,daunorubicin,paclitaxel and mitomycin were detected by MTT assay.Results The cloning efficiency of ABCG2High and ABCG2Low cells was 25.24%?1.18% and 16.28%?1.11%,respectively,on day 7.ABCG2High cells in S phase accounted for 41.5%,while the ABCG2Low cells in the G1/M0 phase went up to as high as 63.9%.The expression of ABCG2 in original ABCG2HighCNE2/DDP cells was 98%,while in ABCG2Low CNE2/DDP cells was 2%.However,at the 5th passage,the expression of ABCG2 decreased to 75% in the former cells and increased to 20% in the latter ones.The expression of ABCG2 mRNA in ABCG2High cells was obviously higher than that in ABCG2Low cells,while there was no difference in expression of other drug resistance genes between two cell lines.The growth velocity of ABCG2High cells exceeded that of ABCG2Low cells in early stage,but it slowed down 7 days later.MTT assay showed that IC50 of ABCG2High cells to 8 kinds of anti-tumor drugs were twofold higher than that of ABCG2Low cells,with statistically significant difference between them(P

Objective:To investigate the inducing effects of sunitinib malate on expression of NKG2D ligands in nasopharyngeal carcinoma cell ABCG2high CNE2/DDP.Methods:ABCG2highCNE2/DDP cells and Allo-NK cells were isolated by magnetic activated cell sorting(MACS).Flow cytometry was used to evaluate the purity of isolated cells and the expression of NKG2D-ligands on target cells before and after incubation with sunitinib malate.Then the cytotoxic sensitivity of treated and un-treated ABCG2high CNE2/DDP cells to Allo-NK cells were measured by LDH releasing assay.Results:The positive rate of ABCG2 in ABCG2highCNE2/DDP cells was(91.40?2.32)%.More than 90% of isolated Allo-NK cells were proven to be CD3-CD16+CD56+ cells.The expression of MICA,MICB,ULBP1,ULBP2 and ULBP3 on ABCG2high CNE2/DDP cells incubated with sunitinib malate increased from(2.92?0.33)%,(4.27?0.33)%,(5.80?0.62)%,(11.10?3.15)%,and(7.75?1.14)% to(89.12?4.56)%,(66.10?2.22)%,(67.56?4.19)%,(69.37?8.83)%,and(63.28?3.31)%,respectively.At the E ∶T ratios of 10 ∶1 and 20 ∶1,the cytotoxic sensitivities of ABCG2high CNE2/DDP cells to Allo-NK cells increased from(15.32?13.86)% and(27.26?6.81)% to(41.12?4.12)% and(57.25?2.37)%,respectively,after treatment with sunitinib malate,with significantly difference found in the cytotoxic sensitivities of target cells in each group before and after sunitinib malate treatment(F=15.58,P=0.000).Conclusion:Sunitinib malate can up-regulate expression of NKG2D-ligands(MICA/B,ULBP1-3)in ABCG2high nasopharyngeal carcinoma cells,which results in higher cytotoxic sensitivity to Allo-NK cells.

Interferon and nucleotide/nucleoside analogues are currently widely used in the management of chronic hepatitis B virus ( HBV) infections, but are facing problems such as poor sustained virologic response, low HBsAg clearance rate and a high risk of recurrence after drug withdraw.Exploring new target of anti-HBV agent has become a hot topic in recent years.This paper reviews the research progress on new anti-HBV targets and related drugs.

Objective To investigate the association of restriction fragment length polymorphisms (RFLP) in p.S267F of SLC10A1 gene with clinical outcomes of hepatitis B virus (HBV) infection. Methods Clinical data of 1 268 patients with HBV infections admitted in Public Health Clinical Center Affiliated to Fudan University during July 2014 and February 2015 were collected.Polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP) method was used to genotype the p .S267F of SLC10A1 gene in all patients, and the potential association between variants in p .S267F of SLC10A1 gene and the clinical outcomes of HBV infection was analyzed .Results Among 1 268 patients with HBV infections, 1 226 were of genotype CC, and 42 were of genotype CT, so the variation rate in p.S267F was 3.31%(42/1 268).Compared with patients with genotype CC , patients with genotype CT had a higher incidence of acute HBV infections (13.6%vs.28.6%,χ2 =19.819, P<0.05) and a lower incidence of HBV-related liver cirrhosis or hepatocellular carcinoma (13.9% vs.4.8%, χ2 =18.945, P <0.05). Conclusion RFLP in p.S267F of SLC10A1 gene may be associated with chronicity and aggravation of HBV infection, and genotype CT is possibly a protective factor .

Objective To construct hantavirus Z37 envelope glycoprotein genes G1 and G2 recombinant and express in eukaryocyte. Methods Using three pairs of primers based on hantavirus Z37 M gene sequences. PCR products F1, F2 and G2 were obtained by PCR from the Plasmid pGEMZ37(containing partial cDNA of G1) and Plasmid pCUMZ37(containing partial cDNA of G1 and whole cDNA of G2). G1 PCR products were also obtained by fusing F1 and F2 PCR products. The G1 and G2 PCR products were then digested by BamHⅠ and XhoⅠ and cloned into the corresponding sites of expression vector pcDNA3.1 respectively. The recombinants pcDNA3.1 G1 and pcDNA3.1 G2 were identified by digestion with endonuclease BamHⅠ and XhoⅠ and confirmed by sequencing. After transfecting COS 7 cells by Calcium phosphate/DNA precipitating method, indirect immunofluorescence assay (IFA) was used to verify the transient expression of HV Z37 G1 or G2 protein in COS 7 cells. Results The recombinant expression plasmids pcDNA3.1 G1 and pcDNA3.1 G2 were constructed. After transfection with the above recombinant expression plasmids, specific antigens were detected within cells by IFA. Conclusions The reombinant expression plasmids pcDNA3.1 G1、pcDNA3.1 G2 were constructed successfully and expressed transiently in eukaryocyte.

Objective To examine whether hantavirus Z37 envelope glycoprotein genes G1 and G2 recombinants could elicit humoral immune responses in mice. Methods We vaccinated BALB/C mice with plasmid pcDNA3.1 G1 and pcDNA3.1 G2 respectively by intramuscular and gene gun inoculation. Serum anti hantavirus antibodies were detected by IFA and neutralizing antibodies (NAb) were detected by plaque reduction neutralization test with immunoperoxidase staining. The mice given intramuscular inoculation were divided into 3 groups, each group containing 5 mice: groupⅠas control inoculated with 100 ?g pcDNA3.1 ; groupⅡ with 100 ?g pcDNA3.1 G1; group Ⅲ with 100 ?g pcDNA3.1 G2. Each mouse was immunized three times. 4 weeks after primary vaccination, 2 boosts were given at 3 week intervals. The mice given gene gun inoculation were divided into 3 groups, each group containing 3 mice: groupⅠ as control with 1.5 ?g pcDNA3.1; group Ⅱ with 1.5 ?g pcDNA3.1 G1; group Ⅲ with 1.5 ?g pcDNA3.1 G2.Each mouse was immunized two times. 4 weeks after primary vaccination, 1 boost was given. Results 1. In mice given intramuscular inoculation, serum anti hantavirus antibodies were detected 3 of 5 in group Ⅱ, 2 of 5 in group Ⅲ.IFA titers 1∶20～1∶80. NAb were detected 1 of 5 in group Ⅱ, 2 of 5 in group Ⅲ. NAb titers 1∶10～1∶20. 2.In mice given gene gun inoculation, serum anti hantavirus antibodies and NAb were detected in all experimental group mice, IFA titers 1∶20～1∶320, NAb titers≥1∶10. Conclusions Genetic immunization with HV Z37envelope glycoprotein genes G1 and G2 recombinants could elicit effective humoral immune response in mice.