OBJECTIVETo study the inhibition of berberine (BBR) against ECV-304 apoptosis induced by Staphylococcus aureus (S. aureus).

METHODSECV-304 cells were pre-treated with 128 microg/mL BBR for 2 h and then S. aureus was added (1:100). The viability of cells was detected by MTT (3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological changes were observed by Hoechst 33258 staining. The protection of BBR for infected cells was detected by DNA Ladder.

RESULTSECV-304 cells' viability were not obviously affected by berberine. But S. aureus induced ECV-304 cells' viability could be significantly inhibited by pre-treatment of BBR (P < 0.05). Besides S. aureus-induced ECV-304 apoptosis could be reduced, with significantly lessened apoptotic body and unobvious DNA degradation.

CONCLUSIONBBR could significantly inhibit S. aureus induced ECV-304 apoptosis.

Apoptosis ; drug effects ; Berberine ; pharmacology ; Cell Line ; Human Umbilical Vein Endothelial Cells ; drug effects ; microbiology ; pathology ; Humans ; Staphylococcus aureus

Objective To analyze the epidemiological characteristics of Mycobacterium tuberculosis by enterbaeterial repetitive intergenic consensus-polymerase chain reaction(ERIC-PCR)DNA fingerprint. Methods Mycobacterium tuberculosis positive sputum samples between September 2003 to May 2006 were collected and cultured.Chromosomal DNA were extracted and ERIC-PCR DNA fingerprinting was analyzed by software,such as RAPD PHYLIP and Treeview.Results A total of 42 different fingerprints were detected.Phylogenetic analysis showed that they could be classified into three clusters,the clustering rate was 72.6%.The characteristics of ERIC-PCR fingerprint patterns were related to age,drug resistance,and type of resistance.Conclusions ERIC-PCR DNA fingerprinting technique used in this study is good for epidemiological studies with its strong discrimination,simplicity and rapidness.A high level of recent transmission is found in our city.

Objective:To investigate the molecular interaction between non-structural protein 3 serine protease of hepatitis C virus(HCV)and wild type P53,and to lay the basis for elucidating the mechanism of oncogenesis of hepatocellular carcinoma(HCC)after infection of HCV.Methods:The recombinant plasmids,pGAD424-NS3,pGAD424-NS315aa- and pGAD424-NS330aa-,were constructed and the interaction between NS3 serine protease and its cofactor NS4A,the interaction between wild type P53 and NS3 serine protease and its N-truncated mutants were dectected qualitatively and quantitatively in yeast two-hybrid system.Results:The results indicated that interaction existed not only between full-length NS3 serine protease and P53,but also between N-truncated mutants of NS3 serine protease and P53.Furthermore,the difference between enzyme activity unit(IU)of β-gal induced by these interactions was not significant(P>0.05).Conclusions:NS3 serine protease of hepatitis C virus and its N-truncated mutants can interact with wild type P53,and the region of NS3 serine protease involved in the interaction may be located in its C-terminal,but not in its N-terminal.

OBJECTIVETo evaluate the different effect on the expression of Calcitonin gene related peptide (CGRP)and neuropeptide Y (NPY) between tissue engineered bone with vascular bundle graft in vivo and that with sensory nerve tract graft in vivo.

METHODThirty-six healthy New Zealand rabbits were divided into 3 groups randomly and equally: vascular bundle group (A), sensory nerve tract group (B), tissue-engineering group (C). Group A segmental bone defect of 1.5 cm long was made at the right femur in each animal. After plate fixation, the defects were implanted respectively with the engineered bone prepared in the above-mentioned 3 methods. At 3, 6 and 12 months post-operatively, the distribution of CGRP and NPY in the new bone were detected by immunohistochemistry and analyzed semi-quantitatively by image analysis software.

RESULTSCGRP and NPY immuno-histochemical results indicated their contents increased significantly in all 3 groups as time passed (P = 0.000). Compared with group B, the contents of CGRP and NPY in group A significantly increased at 3 months (P = 0.000), but there was no statistic difference between them at 6 or 12 months (P > 0.05). The expression of CGRP and NPY in both group A and B were significantly more than that in group C at 3, 6 or 12 months (P = 0.000).

CONCLUSIONImplantation of vascular bundle into tissue-engineered bone can significantly improve the CGRP and NPY contents at early 3 months comparing with Implantation of sensory tract into tissue-engineered bone, but the changes are not significant at 6 or 12 months post-operatively.

Animals ; Blood Vessels ; transplantation ; Bone Substitutes ; Calcitonin Gene-Related Peptide ; metabolism ; Disease Models, Animal ; Femur ; injuries ; Male ; Neuropeptide Y ; metabolism ; Peripheral Nerves ; transplantation ; Rabbits ; Random Allocation ; Tissue Engineering

OBJECTIVETo evaluate the effect and possibility of surgical ablation of the pulmonary vein orifices under direct vision with transballoon ultrasound ablation catheter for patients with permanent atrial fibrillation and rheumatic valve disease.

METHODS21 consecutive patients with rheumatic valve disease and permanent atrial fibrillation undergoing mitral valve replacement surgery were enrolled for this study from December 2002 to September 2003. All the cases were divided into 2 groups by whether or not receiving an additive pulmonary vein ablation procedure. The test group [6 male, 5 female, aged (51.55 +/- 7.83) years, atrial fibrillation duration (5.50 +/- 5.40) years, left atrial diameter (7.27 +/- 1.39) cm, LVEF (53.95 +/- 4.54)% and NYHA class II - IV] undertook a surgical isolation of the pulmonary vein orifices by using a transballoon ultrasound ablation catheter addition to routine mitral valve replacement. The control group [3 male, 7 female, aged (53.30 +/- 7.86) years, atrial fibrillation duration (4.50 +/- 3.47) years, left atrial diameter (6.74 +/- 0.62) cm, LVEF (56.91 +/- 3.78)% and NYHA class II - IV] received the valve replacement surgery alone.

RESULTSThere were not any complications in both groups. With an electrical cardioversion 3 months after the surgery, 73% patients in the ultrasound ablation group were free from AF over 1 year while only 10% patients in control group (P=0.003). During an average follow-up duration of (45.92 +/- 4.61) months, 63.6% were in sinus rhythm in ultrasound ablation group while none in the control group. Left atrial volume decreased significantly at 1 year after surgery compared to that at 3 months after surgery in the test group [(97.83 +/- 32.39) cm(3) vs. (150.78 +/- 52.32) cm(3), P<0.05], and the end systolic diameter (LAESD) and end diastolic diameter (LAEDD) also decreased [(4.12 +/- 0.39) cm vs. (5.09 +/- 0.98) cm, P<0.05, respectively], while there were no apparently changes in the control group.

CONCLUSIONSAblation of the orifices of the pulmonary veins under direct vision with transballoon ultrasound ablation catheter during mitral valve surgery seems effective to maintain sinus rhythm after electrical cardioversion and could be performed safely. The function of left atrial and cardiac output improved during long term follow-up of 46 months.

Atrial Fibrillation ; therapy ; Catheter Ablation ; methods ; Catheterization ; Female ; Heart Valve Diseases ; therapy ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; Rheumatic Heart Disease ; therapy ; Ultrasonic Therapy

OBJECTIVETo investigate the effect of rabbit saphenous and sciatic nerve homogenates on the proliferation and calcification of rabbit osteoblasts in vitro.

METHODThe saphenous nerves (sensory nerves) and the muscular branches of the sciatic nerve (motor nerve) were collected from 48 New Zealand white rabbits to prepare the nerve tissue homogenates. Bone marrow mesenchymal stem cells (MSCs) were isolated from the rabbits and cultured in vitro, and after 14 days of routine osteogenic induction, the resultant osteoblasts were identified by immunohistochemistry, alkaline phosphatase (ALP) and Alizarin red S staining. The osteoblasts were then incubated in the induction medium containing the saphenous (sensory nerve group) or sciatic homogenates (motor nerve group), with the cells in the dexamethasone-containing, dexamethasone-free osteogenic induction medium and control medium as the control. The proliferation, total protein and ALP activity of the osteoblasts were measured every other day until the 8th day, and Alizarin red S staining was used for quantitative analysis of calcification of the cells after two weeks.

RESULTSThe application of the saphenous nerve homogenates significantly promoted cell proliferation, total protein and ALP activity (P<0.01, P<0.05 and P<0.05), while exposure of the osteoblasts to dexamethasone inhibited the cell proliferation (P<0.001). Compared to dexamethasone-free group, the saphenous homogenates enhanced the mineralization of the osteoblasts (P<0.001).

CONCLUSIONSaphenous nerve homogenates significantly promotes the proliferation, differentiation, ALP activity and mineralization of rabbit osteoblasts, but sciatic nerve homogenates do not show osteogenic effects on the cells.

Animals ; Bone Marrow Cells ; cytology ; Calcification, Physiologic ; drug effects ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media ; pharmacology ; Female ; Male ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; physiology ; Rabbits ; Sciatic Nerve ; chemistry ; Sensory Receptor Cells ; chemistry ; Tissue Extracts ; pharmacology

OBJECTIVETo explore the risk factors of acute lymphoblastic leukemia (ALL) recurrence in adult patients and establish a prognosis index (PI) calculation model in order to improve the prevention strategy of ALL in adults.

METHODS104 adult ALL patients from Blood Diseases Hospital & Chinese Academy of Medical Sciences between August 2008 and November 2011 were enrolled. COX proportional hazards regression stratified by Dummy variable was used to set up the prediction model; Kaplan-Meier method and Log-rank test were used to estimate and compare the survival. After calculated individual PI value, patients' expected survival should be estimated by groups.

RESULTSThe overall median survival of adult ALL patients was 22.00 months (95% CI 17.00-27.00). COX regression analysis showed that chemotherapy group patients had a higher risk of recurrence than of ASCT group while setting treatment as the dummy variable (RR=2.052, 95%CI 0.877-4.799, P=0.007). Stratified Analysis showed that the risk factors of B-ALL recurrence in adult patients included HGB <100 g/L (RR=0.186, 95% CI 0.068-0.512, P=0.001), CNSL (RR=7.767,95% CI 2.951- 20.433, P=0.001), number of consolidation chemotherapy<3 (RR=0.445, 95% CI 0.211-0.940, P=0.034) and Ph chromosome positive (RR=2.771, 95% CI 1.353-5.674, P=0.005). Grouped by the PI value, the expected survival of each individual patient could be estimated as PI=0.58 base.

CONCLUSIONHGB, CNSL, number of consolidation chemotherapy and Ph chromosome were independent risk factors of B-ALL recurrence in adult patients. PI value could predict the survival of adult ALL patients and provide reference for individual therapy and prognostic evaluation.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; epidemiology ; pathology ; Prognosis ; Recurrence ; Risk Assessment ; Risk Factors ; Young Adult

Puerarin has the anti-Alzheimer's disease (AD) activity,which can reverse nerve injury induced by Aβand inhibit neuronal apoptosis.However,its potential pharmacodynamic mechanism still needs to be further researched.The occurrence and development of AD is due to the change of multiple metabolic links in the body,which leads to the destruction of balance.Puerarin may act on multiple targets and multiple metabolic processes to achieve therapeutic purposes.Quantitative proteomic analysis provides a new choice to understand the mechanism as completely as possible.This research adopted SH-SY5Y cells induced by Aβ_(1-42)to establish AD cell model,and Aβimmunofluorescence detection showed that Aβdecreased significantly after puerarin intervention.The mechanism of puerarin reversing SH-SY5Y cell injured by Aβ_(1-42)was further explored by using label-free non-labeled quantitative technology and Western blot detection based on bioinformatics analysis result.The results showed that most of the differential proteins were related to biological processes such as cellular component organization or biogenesis,cellular component organization and cellular component biogenesis,and they mainly participated in the top ten pathways of P value such as pathogenic Escherichia coli infection,m TOR signaling pathway,regulation of autophagy,regulation of actin cytoskeleton,spliceosome,hepatocellular carcinoma,tight junction,non-small cell lung cancer,apoptosis and gap junction.Annexin V/PI flow cytometry and TUNEL were used to detect apoptosis,and the results showed that Aβdecreased significantly and the rate of apoptosis decreased significantly after puerarin intervention.Western blot analysis found that the protein expression level of autophagy related protein LC3Ⅱwas up-regulated after Aβinduction,and the degree of this up-regulation was further enhanced in puerarin intervention group.The trend of the ratio of LC3Ⅱ/LC3Ⅰamong groups was the same as the protein expression level of LC3Ⅱ，the protein expression level of p62 in the control group,AD model group and puerarin intervention group decreased successively.Protein interaction network analysis showed that CAP1 was correlated with TUBA1B,HSP90AB2P,DNM1L,TUBA1A and ERK1/2,and the correlation between CAP1 and ERK1/2 was the highest among them.Western blot showed that the expressions of p-ERK1/2,Bax and CAP1 were significantly down-regulated and the protein expression level of Bcl-2 was significantly up-regulated after puerarin intervention.Therefore,puerarin might improve the SH-SY5Y cells injured by Aβ_(1-42)through the interaction of multiple biological processes and pathways in cells multiple locations,and CAP1 might play an important role among them.
Amyloid beta-Peptides ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; Cell Line, Tumor ; Humans ; Isoflavones/pharmacology* ; Lung Neoplasms ; Proteomics

Objective: Early T-cell precursor acute lymphoblastic leukemia (ETP-ALL) is a recently recognized high-risk T lymphoblastic leukemia subgroup. The optimal therapeutic approaches to adult patients with ETP-ALL are poorly characterized. In this study, we explore the efficacy and outcome of allogeneic hematopoietic stem cell transplantation (allo-HSCT) for ETP-ALL. Methods: The clinical data of 23 patients with ETP-ALL receiving allo-HSCT from 2010 to 2018 were retrospectively analyzed. Patients with ETP-ALL were diagnosed based on the characteristic immunophenotypes. Second-generation sequencing was done in all patients. As to the donors, 12 patients had haploidentical donors (Haplo-HSCT) , 7 HLA-matched sibling donors (Sib-HSCT) and 4 HLA-matched unrelated donors (URD-HSCT) . Before transplantation, 19 patients achieved complete remission (CR) and 4 patients without. Results: The main clinical features of ETP-ALL included high white blood cell counts in 5 patients, splenomegaly in 14, lymphadenopathy in 19, and thymus masses in 5. According to cytogenetic and molecular characteristics, 11 patients had gene mutations related to myeloid tumors, and 7 with high risk Karyotype. After first induction regimen, 14/23 patients achieved CR. 5 patients reached CR after more than 2 cycles of chemotherapy, while another 4 patients did not reach CR. After allo-HSCT, 22 patients were successfully implanted. The median time of granulocyte and platelet reconstitution was +12 and +19 days. One patient died of transplant-related infection at +14 days. The estimated 18-month overall survival (OS) and relapse-free survival (RFS) rates were (55.0±14.4) % and (48.1±14.7) % respectively. Transplant-related mortality was 4.3%. The median OS in patients achieving CR before transplantation was 20 months, however, that in patients without CR was only 13 months. OS and RFS between haplo-HSCT and sib-HSCT were comparable (P=0.460 and 0.420 respectively) . Conclusions: Allo-HSCT is an effective therapy in some patients with ETP-ALL. Salvage HSCT cannot overcome the poor outcome. Haplo-HSCT and sib-HSCT in ETP-ALL patients have the similar clinical outcome.
Adult ; Hematopoietic Stem Cell Transplantation ; Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Precursor Cells, T-Lymphoid ; Remission Induction ; Retrospective Studies