Objective To clone and identify human genes transactivated by homo sapiens HCV FTP2 by constructing a cDNA subtractive library with suppression subtractive hybridization tech- nique.Methods Suppression subtractive hybridization(SSH)and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV FTP2.The mRNA was iso- lated from HepG2 cells transfected pcDNA3.1(-)-HCV FTP2 and pcDNA3.1(-)empty vector re- spectively,and SSH method was employed to analyze the differentially expressed DNA sequence be- tween the two groups.After digestion with restriction enzyme Rsa I,small-size cDNAs were ob- tained.Then tester cDNA was divided into two groups and ligated to the specific adaptor I and adap- tor 2 respectively.After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5?.Futhermore,the cDNA was se- quenced and analyzed in GenBank with Blast search after PCR.Results The subtractive library of genes transactivated by HCV FTP2 was constructed successfully.The amplified library contains 71 positive clones.Colony PCR shows that 56 clones contain 200～1000 hp inserts.Sequence analysis was performed in 24 clones randomly,and the full length sequences were obtained with bioinformatics method.Altogether 20 coding sequences in total were obtained,consisting of 19 known and 1 un- known.Conclusion The obtained sequences may be target genes transactivated by HCV FTP2,and some genes coding proteins involved in cell cycle regulation,metabolism and cell apoptosis.

0.05).Conclusion Levofloxacin treating community-acquired pneumonia than gatifloxacin have more medicine economics advantage and it is in the prior choice.

0.05).Conclusion Gatifloxation is safe and efficient in the treatment of community acquired pneomonia.

Objective To screen promoter binding protein of cyclin B2 by using human liver cDNA library, and investigate the expression and regulation mechanism of cyclin B2 gene. Methods By using cyclin B2 biotinylated promoter DNA as the selective molecule, the T7select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment, and the DNA fragment was cloned into pGEM-Teasy vector. Results Sequence analysis was performed in 20 positive plaques, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 6 coding sequences were obtained, all of which were known ones. Conclusion Cyclin B2 promoter binding proteins were screened. The results will be useful for further study the expression and regulation mechanism of cyclin B2.

Ascites ; complications ; etiology ; Heart Failure ; complications ; etiology ; Hemochromatosis ; complications ; diagnosis ; Humans ; Middle Aged

Objective To study the changes and significance of the frequencies of circulating follicular helper T cells (cTfh) and circulating regulatory follicular T cells (cTfr) as well as the cTfh/cTfr ratio in neuromyelitis optica spectrum disorder (NMOSD).Methods The frequencies of cTfh,cTfr and B cells in patients with NMOSD and health controls(HCs) were measured by flow cytometry.Enzyme-linked immunosorbent assay was used to detect the level of IL-21 and AQP4-Ab in patients and HCs.Results The frequencies of cTfh and B cells,the cTfh/cTfr ratio and the plasma level of IL-21 werc significantly higher in the relapsing patients than those in the remitting patients and HCs(P ＜ 0.05),and the cTfr level in the relapsing patients was lower than that in the remitting patients and healthy population (P ＜ 0.05).But no statistical differences were observed in the above indexes between the remitting paticnts and HCs.There was also no significant difference in AQP4-Ab level between the patients with relapse and remission (P ＞ 0.05).The frequency of cTfh in the patients wasc positively correlated with the level of B cells and IL-21(P ＜ 0.05),and the frequency of cTfr was negatively correlated with B cells and IL-21 (P ＜ 0.05).The ratio of cTfh/cTfr was positively correlated with B cell frequency and IL-21 level (P ＜ 0.05).AQP4-Ab level had no correlation with the frequencies of cTfh cells and B cells,cTfh/cTfr ratio and IL-21 concentration (P ＞ 0.05).Conclusion The changes in the frequencies of cTfh and cTfr as well as the imbalanced cTfh/cTfr ratio may promote the activation of humoral immunein NMOSD and participate in the pathogenesis of this disease.

Objective To investigate the effect of hyperuricemia(HUA) on inflammation and endothelin-1 (ET-1) production and treatment of Benzbnomanone in hypertensive patients.Methods 90 initial hypertensive patients were enrolled from the inpatient division and clinic of our hospital,60 patients of them were identified HUA,and 30 patients were normal in uric acid as control.All these hypertensive patients with HUA were treated with basic anti-hypertensive drugs,of them 30 patients were additionally treated with Benzbromarone table 50mg for 8 weeks.The levels of inflammation indices and ET-1 were compared between these hypertensive patients with HUA and hypertensive patients with normal serum uric acid,also hypertensive patients with HUA treated with or without Benzbromarone for 8 weeks.Results Compared with the hypertensive patients with normal serum uric acid,levels of ET-1,interleukin-1 (IL-1) and high-sensitive C reactive protein (hsCRP) were higher in the hypertensive patients with HUA.Also,the levels of these indices were positively correlated with the level of serum uric acid [(86.6 ± 4.8) pg/ml vs (82.4 ±6.9)pg/ml; (47.6 ±6.2)mg/L vs (19.1 ±4.1) mg/L; (3.4 ±0.8)mg/L vs (2.9 ± 1.1)mg/L,r =0.81,0.74,0.83,all P ＜ 0.05].Benzbromarone could effectively decrease the levels of ET-1,IL-1and hsCRP in the hypertensive patients with HUA [(49.8 ± 5.0) pg/ml vs (87.5 ± 5.9) pg/ml ; (17.6 ±8.8) mg/L vs (48.2 ± 7.0) mg/L; (1.7 ± 0.7) mg/L vs (3.5 ± 0.9) mg/L,all P ＜ 0.05].Conclusions HUA could increase the levels of inflammation and ET-1,while Benzbromarone effectivelv decreased these changes.Decreasing the level of serum uric acid would retard the process of atherosclerosis in the hypertensive patients with HUA.

Acute myocardial infarction (AMI) is one of the most common cardiovascular diseases. The clinical pathway is the therapeutic program for disease-specific treatment and its implementation may reduce both the duration and cost of the hospital stay. This study aims to construct and evaluate the efficacy of clinical pathways (CPs) based on integrated traditional Chinese and Western medicine for patients with AMI.

AIM:To investigate the effect of pretreatment of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) on the proliferation and the differentiation of mesenchymal stem cells(MSCs) into cardiomyogenic cells.METHODS:The MSCs,isolated primarily from bone marrow,and purified by passage culture,were obtained from the adult rats of four groups:the rats were pretreated by 5 daily injections of SCF;the rats were pretreated with G-CSF;the rats were pretreated with SCF and G-CSF;the rats were treated without any intervention.The 4th passage of MSCs was labeled by DAPI and cellular cycle analysis was conducted by flow cytometry before co-culture.The neonatal rat cardiomyocytes cultured for 3 days were co-cultured with DAPI-MSCs.The percentage of the differentiation of MSCs into cardiomyogenic cells during the five co-culture days was analyzed.The morphologic changes of MSCs and the proteins expression of cardiac myosin heavy chain(MHC) and troponin T(TnT) were recorded respectively with digital microscope camera system and immunofluorescence technique.The percentage of the differentiation of MSCs into cardiomyogenic cells was also calculated.RESULTS:The percentage of MSCs in G0/G1 phase in SCF/G-CSF group was significantly lower than that in SCF group,G-CSF group and the control group.The percentage of MHC protein-positive MSCs in SCF/G-CSF group was markedly higher than that in SCF group,G-CSF group and the control group,and that in SCF group and G-CSF group was significantly higher than control group.The percentage of TnT protein-positive MSCs in SCF/G-CSF group,SCF group and G-CSF group was significantly higher than that in control group.CONCLUSION:SCF and G-CSF show the ability to stimulate the proliferation of MSCs and induce MSCs to differentiate into cardiomyocytes.The combination of using SCF and G-CSF is more effective than using only SCF or G-CSF.

Objective:To investigate surgical methods and clinical effects of tibial pseudarthrosis due to neurofibromatosis type I (NF1) in adult using Ilizarov technique combined with intramedullary nail.Methods:A total of 12 adult with tibial pseudarthrosis due to NF1 treated by Ilizarov technique combined with intramedullary nail from October 2009 to December 2017 were retrospectively included. There were 6 males and 6 females with an average age of 27±8.3 years (range 17-44). All cases presented severe anterior arch in varus or valgus deformity with shortening in an average of 10.8±3.7 cm (range 5.6-16.5 cm). The in-volved levels were at the middle and lower part of tibia. All cases suffered from more than one treatment with failed surgery. One male patient with 5 times of unsuccessful operations. There were 4 cases with severe limping gait and 8 cases with walking with the help of brace (or single crutch) preoperatively. Regarding the family history, seven cases were inherited by father and 5 cases by mother. During surgery, the part of tibial pseudarthrosis and thickened fibrous tissue surroundings like periosteal were removed. The contracture achilles tendon was elongated in open way, and intramedullary nailing cross the ankle joint was applied when the tibial medullary cavity opened. Further, iliac bone grafting and proximal tibial osteotomy were performed with Ilizarov fixator application finally. The tibia was lengthened at a rate of 0.5-1 mm/d at 7 days postoperatively. The healing rate of pseudarthrosis and the length and alignment of limb were evaluated by X-ray routinely. The lower limb function and complications were assessed by self-made table for lower limb deformity correction and functional reconstruction.Results:All patients were followed up for 31-80 months with an average of 47.6±14.7 months. Bone union of pseudarthrosis in all cases was eventually achieved. The tibia was lengthened 5-12 cm with an average of 8.4±2.5 cm. There were 9 cases underwent second surgeries to promote bone healing and to correct residual deformities. The external fixator was used for 25-37 months with an average of 31.5±3.7 months. There was no complication, including neurovascular injury, severe soft tissue or bone infection affecting the clinical effects. All intramedullary nails were not removed finally. The limb function and gait in full weight bearing in 12 patients recovered at the latest follow-up. The evaluation score was 2.4±0.3 (range 2.1-2.8), of which 9 cases were excellent 3 were good. Thus, the excellent and good rate was 100%.Conclusion:The satisfactory clinical effects, including angular deformity correction, pseudarthrosis healing and short limb lengthening, can be achieved in adult with tibial pseudarthrosis due to NF1 by using the combination method of Ilizarov technique and intramedullary nail. However, the treatment duration could be longer.