Objective To synthesize an inhibitory peptide against serine protease of hepatitis C virus(HCV)and explore its inhibition ability in vitro. Methods Based on the sequence characteristics of four natural substrates of protease in HCV,the gene sequence of an inhibitory peptide against HCV serine protease was designed and directly synthesized by PCR. The segment was subcloned into prokaryotic expression vector pBVIL1,resulting in the construction of the recombinant plasmid pBVIL1/IP,which was then transformed into E. coli HB101 strain. Purified by ion exchange chromatography,the expressed protein was added into an in vitro system,which was comprised of the inhibitory peptide(expressed protein),protease and the substrate,i.e. a NS5A-B fragment,in phosphate buffer. Then SDS-PAGE was performed to test the inhibition effect of the polypeptide. Results The recombinant expression vector pBVIL1/IP containing target gene was successfully constructed. The peptide expressed as inclusion boay was identified by SDS-PAGE. The degradation of protease substrate NS5A-B fragment is inhibited proportionally with the increasing concentrations of the peptide. Conclusion The recombinant peptide shows inhibitory effect on HCV protease in vitro.

Brucellosis ; diagnosis ; drug therapy ; Child ; Child, Preschool ; Female ; Humans ; Male

Aged ; Female ; Humans ; Lymphohistiocytosis, Hemophagocytic ; virology ; Orthobunyavirus ; classification

Objective To study the perioperative nursing care of minimally invasive operation in the neurosurgical treatment of laterial flexure spasmodic torticollis (LaFST).Methods Minimally invasive operation was used to treat LaFST.The relationship of inducing factors and emotional factors with clinical manifestation of LaFST was investigated.To give perioperative psychological nursing,operation nursing and rehabilitation training and instruction.Results 72 cases were recovered (81.8％) among 88 cases.12 cases(13.6％) were markedly effective.4 cases(4.6％) showed progress.Conclusions The minimally invasive surgical treatrnent of selective resection of cervical spasmodic muscles and selective neurotomy of cervical nerve for LaFST is safe and effective.Strengthening of perioperative nursing and postoperative rehabilitation instruction is very important for patients' early recovery.

Objective To assess the impact of HLA-A,HLA-B,HLA-DRB1 matching on the prognosis of hematopoietic stem cell transplantation from unrelated donors.Methods A total of 81 patients with hematological malignancies including leukemia,myelodysplastic syndrome(MDS)and lymphoma who underwent hematopoietic stem cell transplantation from unrelated donors from 2007 to 2012 in our department were included in this retrospective analysis.Patients were classified into HLA match group(n=53)and HLA mismatch group(n=28)according to the HLA high-resolution matching.The overall survival (OS),treatmentrelated mortality(TRM),relapse rate(RR),graft-versus-host disease(GVHD)incidence were analyzed.Results The 81 patients were analyzed with a median follow-up of 11.9 months(0.3 to 57.4 months).The OS (66.0％vs 46.4％,P=0.031)and TRM(17.0％vs 42.9％,P=0.017)were significantly different between the HLA match group and HLA mismatch group,while the RR had no significant difference(14.3％vs 32.1％,P=0.111).Multivariate analysis showed HLA matching was an independent prognostic factor of TRM,but not OS.There's no significant difference of aGVHD(22.9％vs 40.9％,P=0.122)and cGVHD (40.0％vs 46.7％,P=0.655)incidence between the two groups,but the incidence of severe aGVHD in HLA match group were much lower(4.2％vs 25.0％,P=0.005)than HLA mismatch group.Conclusion the high-resolution matching of HLA-A,-B,DRB1 affect OS,TRM and the incidence of severe aGVHD in unrelated hematopoietic stem cell transplantation,but not affect RR,the incidence of aGVHD and cGVHD.

Objective To analyze the antibiotic resistance of Branhemella catarrhalis strains isolated from sputum specimens of patients with lower respiratory tract infections from Linyi, Shandong Province, and to explore the relationship between bro genotypes of the strains and their resistance to antibiotic agents.Methods Sputum specimens were colleted from the patients with lower respiratory tract infections in Linyi People ’ s Hospital from the January 2010 to December 2014.The specimens were inoculated into 4 different disks for bacterial isolation and cultivation.β-lactamase detection and drug sensitivity tests were performed, and PCR coupled with restriction endonuclease analysis was employed for bro genotyping.χ2 test was used to compare drug resistance of strains with different bro genotypes.Results A total of 497 Branhemella catarrhalis strains were isolated in five years, among which 221 strains were isolated in winter.All strains were sensitive to ertapenem and chloramphenicol, and the resistance rates to amoxicillin/clavulanate and cefaclor were low (≤2.8%).The strains were highly resistant to compound sulfamethoxazole, erythromycin and ampicillin (47.6%-89.8%), and there was a trend of increasing resistance rates with the year, but no statistically significant difference was observed ( P >0.05 ) .β-lactamases was positive in 412 strains (82.9%), and all of these strains were positive for bro gene, and the resistances to erythromycin, compound sulfamethoxazole, levofloxacin and ampicillin were higher in bro positive strains than those in bro negative strains (χ2 =12.16, 16.18, 8.41 and 200.00,P<0.05).Among bro positive strains, 391 (94.9%) were of genotype bro-1, 21 (5.1%) were of genotype bro-2, and their resistance to antibiotic agents was not of statistical difference ( P >0.05 ).Conclusions Most of Branhemella catarrhalis clinical isolates are β-lactamase producing strains, and bro-1 is the most common genotype.Strains are highly sensitive to carbapenems, cephalosporins andβ-Lactamaseinhibitors, which can be recommended for the treatment of Branhemella catarrhalis-related respiratory tract infections.

Objective A retrospective analysis of patients with T-cell lymphoma (TCL) received autologous peripheral blood stem cell transplantation (APBSCT) was performed to evaluate the outcome of APBSCT.Methods A total of 22 patients who underwent APBSCT from September 2006 to December 2011 in Ruijin hospital were enrolled in the study,including 6 cases of lymphoblastic lymphoma and 16 of peripheral T-cell lymphoma (8 anaplastic large cell lymphoma, 4 PTCL-u, 1 subcutaneous panniculitis-like T-cell lymphoma, 2 nasal type extranodal NK/T and 1 primary cutaneous T-cell lymphoma). All patients were diagnosed based on the WHO Classification of Tumors of Hematopoietic and Lymphoid Tissues. Conditioning regimens were high-dose chemotherapies alone which include 13 cases with BEAM, 4 with ICE and 5 with CBV. The outcomes of the treatment were evaluated according to the revised International Working Group criteria.Results With a median follow-up of 13.1(1-60) months,the predicted 2-year overall survival (OS) and progression-free survival (PFS) after transplantation were (67.6±11.0) ％ and (71.1±11.1) ％,respectively.A total of 6 patients experienced disease progression and 5 patients eventually died of disease. When all these patients based on the remission status before APBSCT (CR1 vs non-CR1) and chemosensitivity (sensitive vs refractory) were further classified, the PFS rates and OS rates were 100 ％ and 91.7 ％ respectively in CR1 or chemosensitive patients which were significantly higher than patients not in CR1 (42.6 ％ ) or with chemoresistant disease (19.0 ％ ). Conclusion Remission status and chemosensitivity at the time of transplantation significantly affect the outcome of APBSCT for TCL patients, thus it can be recommend to perform APBSCT for patients either in CR1 or early stage when the disease remain sensitive to chemotherapy.

Background and purpose: High-dose chemotherapy followed by autologous stem cell transplantation (auto-HSCT) is considered as the ifrst line treatment for patients with relapse/refractory lymphoma after conventional chemotherapy. However, most of these patients still relapse the second time. The purpose of this study was to analyze the efifcacy of the consolidation chemotherapy after autologous stem cell transplantation (HSCT) refractory/relapse lymphoma in high risk. Methods:A total of 38 patients with relapsed/refractory lymphoma including Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) were included, who were underwent auto-HSCT in our transplan-tation department from Jan. 2010 to Dec. 2013. In treatment group, 19 patients received 2 courses of consolidation che-motherapy after auto-HSCT every 2 to 3 months, with the regimen of mini-BEAM or modiifed mini-CBV. Another 19 patients had no chemotherapy after auto-HSCT as control group. Results:The median follow-up duration was 17.2 and 7.5 months in the treatment and control group respectively. The follow-up data demonstrated prolonged progression-free survival (PFS) in the treatment group than the control group [24.7 months vs 7.8 months, P=0.029 under intend-to-treat analysis ITT;24.7 months vs 5.2 months, P=0.01 under per protocol analysis(pp)]. There is also a trend of improved overall survival (OS) in the treatment group (P=0.055, ITT). Conclusion:Consolidation chemotherapy after auto-HSCT for refractory/relapsed lymphoma patients delay the relapse and tend to improve the overall relapse rate.

Objective To assess the impact of antithymocyte globulin (ATG) on the incidence of graft-vs-host disease (GVHD) in hematopoietic stem cell transplantation from unrelated donors.Methods A total of 92 patients with hematological malignancies including leukemia,myelodysplastic syndrome (MDS) and lymphoma who underwent hematopoietic stem cell transplantation from unrelated donors from January 1999 to December 2011 were included in this retrospective analysis.Patients were classified into ATG group (n =66)and non-ATG group (n =26) according to the GVHD prophylaxis regimen.The incidence of acute GVHD (aGVHD) and chronic GVHD (cGVHD),risk factors of aGVHD and cGVHD and impact of ATG on the overall survival (OS),treatment related mortality (TRM) and relapse rate were analyzed.Results Grade Ⅱ-Ⅳ aGVHD (26.7 ％ vs 44.0 ％,P=0.12) or grade Ⅲ-Ⅳ aGVHD (13.3 ％ vs 8.0 ％,P =0.74) were not significantly different between ATG and non-ATG group.However,the incidence of cGVHD in the ATG group was significantly lower (34.0 ％ vs 72.2 ％,P =0.005) than non-ATG group.The incidence of extensive cGVHD was also significantly reduced (10.0 ％ vs 44.4 ％,P =0.005) compared to non-ATG group.In multivariate analysis,the use of ATG prophylaxis significantly decreased the cGVHD (RR =0.22,95 ％CI 0.081-0.599,P =0.003) while one allele mismatch of human leukocyte antigen (HLA) was associated with increased risk of cGVHD (RR =3.25,95 ％ CI 1.39-7.61,P =0.007).As to the extensive cGVHD,the use of ATG was the only independent factor (RR =0.05,95 ％ CI 0.009-0.240,P ＜ 0.001).With a median follow-up of 12 months (1-84 months),ATG prophylaxis had no impact on OS rate (60.4 ％ vs 43.1％,P =0.41),TRM rate (19.8 ％ vs 34.3 ％,P =0.43) and relapse rate (40.6 ％ vs 33.6 ％,P=0.54).Conclusion In hematopoietic stem cell transplantation from unrelated donors,ATG prophylaxis total dose of 6 mg/kg may significantly decrease the incidence of cGVHD and extensive cGVHD without increase of TRMand relapse rate and impairment of OS.

Tetrahydrobiopterin (BH4) is an essential cofactor for all three nitric oxide synthase (NOS isoforms), which plays an important role in vascular diseases. GTP cyclohydrolase 1 (GCH 1) is the first-step and rate-limiting enzyme for BH4 biosynthesis in its de novo pathway. Common GCH1 gene variant C+243T in the 3'-untranslated region predicts NO excretion. The present study examined the predictive role of GCH 1 gene 3'-UTR C+243T variant in the long-term outcome of ischemic stroke. A total of 142 patients with first-onset ischemic stroke were recruited and detected for genotype of GCH1 3'-UTR C+243T by a TaqMan SNP Genotyping assay. Subsequent vascular events and death were determined over a 5-year follow-up period. The frequency of GCH1 3'-UTR +243 C/T or T/T genotype was significantly increased in patients with endpoint events as compared with those without events (74% vs 57.8%, P=0.06). Cox regression survival analysis indicated that an increased probability of death or new vascular events was found in patients with GCH1 3'-UTR +243 C/T or T/T genotype compared with those with GCH1 3'-UTR C/C genotype (40.6% vs 25.5%), GCH1 3'-UTR +243 C/T or T/T genotype relative to GCH1 3'-UTR C/C genotype was associated with the increased risk of death or vascular events even after adjustment for other risk factors (OR=2.171, 95% CI: 1.066-4.424, P=0.033). It was concluded that GCH1 3'-UTR C+243T variant was an independent predictor of worsening long-term outcomes in patients with first-onset ischemic stroke.