Objective To observe the therapeutic efficacy of Treating chronic hepatitis B with compound BieJiaRuanGan pill plus entacavir. Methods 168 patients were randomly recruited into a treatment group and a control group. The treatment group was treated with compound BieJiaRuanGan pill plus entacavir and the control group was treated with entacavir exclusively, with a course of 48 weeks. Results Both groups showed statistical significance in improving the liver function, peripheral T lymphocyte subsets and indexes of serum liver fibrosis after the treatment (P<0.01 or P<0.05 ). The therapeutic effects in the treatment group were significant better than the control group after the treatment (P<0.01orP< 0.05 ) . Conclusion The treatment of chronic hepatitis B with compound BieJiaRuanGan pill plus entacavir is better than using entacavir exclusively.

The 1.23 kb DNA fragment encoding the early protein EP0 of pseudorabies virus (PRV) Ea strain was amplified by PCR technique and cloned into pBluescriptII sk+.Three sequencing plasmids containing the partial fragment of the EP0 gene were constructed and the sequences were obtained by Sanger's sequencing technique. Compared with PRV InFh strain, there were multipile site-mutations and a deleted-mutation in the EP0 gene of PRV strain Ea，and the diversity of amino acid residues also existed.Then, the EP0 gene was inserted into an expression vector, pET-28a, fused into the downstream of the 6ΧHis-Tag in frame, to yield the expression plasmid pETEP0. After induction by IPTG, a high expression of fusion protein was obtained, SDS-PAGE analysis and Western blotting showed that the fusion protein was 62kD and the protein was specific to antisera against PRV Ea strain. This indicated that the EP0 gene be expressed in BL21(DE3) and the expression products have immuno-genicity.

This study was aimed to explore the effects of insulin on expression of insulin receptor (IR) and insulin-like growth factor-I receptor (IGF-IR) in Reh cells and promoting effect on proliferation of Reh cells. The proliferation of Reh cells were evaluated by CCK-8 assay. The expression levels of IR and IGF-IR mRNA in Reh cells at different times were detected by real-time quantitative polymerase chain reaction. The results showed that insulin promoted the proliferation of Reh cells in dose- and time-dependent manners. Compared with the control group, insulin promoted the proliferation of Reh cells obviously (P < 0.05). When Reh cells were treated with insulin 10(-9) mol/L for 24, 48 and 72 h, the relative quantity of IR expression (2(-ΔCt1)/2(-ΔCt2)) was 2.2520 ± 0.7431, 1.9956 ± 0.9692 and 3.9766 ± 1.3189, respectively, the relative quantity of IGF-IR expression was 1.0803 ± 0.2238, 1.6026 ± 0.6158 and 3.1013 ± 0.1008, respectively, compared with the control group. The expression levels of IR and IGF-IR mRNA in Reh cells treated with insulin were obviously increased compared with the control group. It is concluded that insulin promotes the proliferation of Reh cells. The high expression levels of IR and IGF-IR may closely related with the growth of leukemia cells.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Insulin ; pharmacology ; Leukemia, Lymphoid ; metabolism ; pathology ; Receptor, IGF Type 1 ; metabolism ; Receptor, Insulin ; metabolism

Objective To observe the characteristics of acute graft-versus-host disease (aGVHD) confined solely to gastrointestinal tract after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Clinical data of 307 patients who received allo-HSCT in our hospital between November 2007 and December 2008 were retrospectively analyzed. Clinical features, risk factors, therap(e)utic effects and prognosis of 38 patients whose aGVHD confined solely to gastrointestinal tract after allo-HSCT were analyzed.Results The incidence of aGVHD was 56. 7% (174/307). aGVHD confined solely to gastrointestinal tract occurred in 38 out of 174 patients (21.8%). The incidence of aGVHD confined solely to gastrointestinal tract was affected by human leukocyte antigen (HLA)-compatible level. The patients transplanted from HLA-matched donor had a higher probability of aGVHD confined solely to the gastrointestinal tract. After treatment with glucocorticoids or anti-CD25 monoclonal antibody, 2 patients who had aGVHD confined solely to the gastrointestinal tract died of uncontrolled aGVHD and concurrent infection. The total effective rate was 94.7%. But for patients who had aGVHD between Ⅱ-Ⅳ degree and not confined solely to gastrointestinal,therapeutic effective rate was 85.2%. There was a difference in therapeutic effect between the two groups (P =0. 015). Anti-CD25 monoclonal antibody was used in 12 patients who had aGVHD confined solely to the gastrointestinal tract and all patients' conditions of aGVHD were controlled. Conclusions aGVHD confined solely to gastrointestinal tract after HSCT was not rare. The incidence of aGVHD confined solely to gastrointestinal tract was higher in HLA-matched sibling donor than in mismatched transplantation.Therapeutic efficacy for the aGVHD confined solely to gastrointestinal tract was good. The favorable outcome might be related with the application of anti-CD25 monoclonal antibody.

OBJECTIVETo study the therapeutic effect of agonistic CD40 monoclonal antibody combined with tumor specific cytotoxic T lymphocyte (CTL) on B lymphoma.

METHODSHuman B lymphoma cell line, Daudi cells, were cultured with CD40 mAb (5C11) for 24 and 48 hours, respectively. Annexin V/PI-binding assay was employed to analyze apoptosis, and FCM to analyze Fas (CD95) expression. Human peripheral monocyte-derived DC were loaded with apoptotic Daudi cells and stimulated by SC11 for further maturation. Tumor specific CTL were generated in vitro by co-culture of mature DC with autologous T lymphocytes. DNA fragmentations of Daudi cells treated with 5C11, CTL or 5C11 combined with CTL were determined by JAM assay. To establish the B lymphoma model, Daudi cells were subcutaneously injected into humanized SCID mice (hu-SCID). 1 or 3 weeks after tumor transfer. tumor-bearing mice were respectively treated with SC11, CTL, 5C11 combined with CTL by intraperitoneal injection. Tumor volume in differently treated mice was measured every week after therapy, and the survival of tumor-bearing mice was recorded.

RESULTS5C11 significantly up-regulated FAS expression in Daudi cells, but had no significant effect on apoptosis rate of Daudi cells. Tumor-specific CTL could effectively kill Daudi cells. Fragmentation of Daudi cells co-cultured with CTL was remarkably enhanced by combination with SC11. Tumor growth in hu-SCID mice was apparently delayed by treatment with SC11, CTL, or SC11 combined with CTL. Moreover, minimal tumor burden mice got 30.0% or 70.0% complete remission (CR), respectively, when received CTL treatment or combination treatment of SC11 with CTL, and the lifespan of tumor bearing mice was also prolonged significantly.

CONCLUSIONSC11 may enhance the sensitivity of Daudi cells to apoptosis by up-regulation of Fas expression and promote cytotoxicity of CTL in vitro and therapeutic effect in vivo.

Animals ; Antibodies, Monoclonal ; immunology ; therapeutic use ; Apoptosis ; immunology ; CD40 Antigens ; immunology ; Cell Line, Tumor ; Coculture Techniques ; Female ; Flow Cytometry ; Humans ; Immunotherapy, Adoptive ; methods ; Lymphoma, B-Cell ; immunology ; pathology ; therapy ; Mice ; Mice, SCID ; Remission Induction ; Survival Analysis ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; Xenograft Model Antitumor Assays ; fas Receptor ; immunology

OBJECTIVE Lapachol is a natural naphthoquinone compound that possesses extensive biological activities. The aim of this study is to investigate the inhibitory effects of lapachol on rat C6 glioma both in vitro and in vivo, as well as the potential mechanisms. METHODS The antitumor effect of lapachol was firstly evaluated in the C6 glioma model in Wistar rats. The effects of lapachol on C6 cell proliferation, apoptosis and DNA damage were detected by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS)/ phenazinemethosulfate (PMS) assay, hoechst 33358 staining, annexinⅤ-FITC/PI staining, and comet assay. Effects of lapachol on topoisomerase I (TOP I) and topoi?somerase Ⅱ (TOP Ⅱ) activities were detected by TOP Ⅰ and TOP Ⅱ mediated supercoiled pBR322 DNA relaxation assays and molecular docking. TOPⅠ and TOPⅡ expression levels in C6 cells were also determined. RESULTS High dose lapachol showed significant inhibitory effect on the C6 glioma in Wistar rats (P<0.05). It was showed that lapachol could inhibit proliferation, induce apoptosis and DNA damage of C6 cells in dose dependent manners. Lapachol could inhibit the activities of both TOPⅠ and Ⅱ. Lapachol-TOPⅠ showed relatively stronger interaction than that of lapachol-TOPⅡ in molecular docking study. Also, lapachol could inhibit TOPⅡ expression levels, but not TOPⅠ expression levels. CONCLUSION These results showed that lapachol could significantly inhibit C6 glioma both in vivo and in vitro, which might be related with inhibiting TOPⅠ and TOPⅡ activities, as well as TOPⅡ expression.

OBJECTIVETo investigate the prevalence of influenza virus infections in children in 2006 using the real-time PCR method.

METHODS(1) Consulting the most conserved sequence NP gene of influenza virus, after comparing with the NP gene sequences of influenza virus in GenBank, one pair of specific primers and one TaqMan probe were designed for each subtype of influenza virus by the software Primer Express. The sensitivity of influenza was evaluated by testing known positive samples which had been two-fold diluted. The specificity of real-time PCR for influenza virus detection was assessed by cross testing 60 isolates of influenza A, 16 isolates of influenza B, and by testing a variety of other respiratory viruses positive samples; (2) 281 nasopharyngeal aspirate samples were detected by real-time PCR and virus isolation; (3) the 12 301 specimens from the patients of Guangzhou Children's Hospital were tested by using the real-time PCR method. Furthermore, the real-time PCR reagent was evaluated by comparing with the result of virus isolation.

RESULTS(1) The sensitivity of real-time PCR developed in this study for influenza A detection was 1:2(22) and for influenza B was 1:2(20) in two-fold serially diluted way. (2) No positive results were found in cross testing of other viruses positive specimens. (3) Influenza virus was detected from 1687 cases (13.71%) out of the 12 301 cases, including 773 cases (45.8%) positive for subtype A and 914 cases (54.2%) positive for subtype B; 455 out of 525 (86.7%) of influenza B positive specimens and 70 out of 525 (13.3%) of influenza A (H1N1) positive specimens were from patients seen during January to April; 419 out of 1118 (37.5%) specimens positive for influenza B and 699 out of 1118 (62.5%) specimens positive for influenza A (H1N1) were from patients seen from May to August. Influenza virus could be identified from 1380 samples by the methods of virus isolation, accounting for 81.80% of the 1687 positive samples detected by real-time PCR. All the influenza virus subtype A was H1N1.

CONCLUSIONThe real-time PCR method developed in this study was sensitive and specific for detecting influenza A and B in clinical specimens. During 2006, influenza A and influenza B co-circulated. The predominant virus was influenza B from January to April, peaking in April. Influenza A (H1N1) prevailed from May to August, with the peak in June.

Child ; China ; epidemiology ; Epidemics ; Humans ; Influenza A Virus, H1N1 Subtype ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Polymerase Chain Reaction ; methods ; Prevalence ; RNA, Viral ; isolation & purification ; Sensitivity and Specificity

OBJECTIVETo review the long-term clinical effect of composite transplantation of allogeneic acellular dermal matrix (ADM) and split thickness skin autograft (STSG).

METHODSNineteen patients with 34 wounds transplanted with allogeneic ADM combined with STSG who were hospitalized from March 2001 to October 2008 were enrolled as composite transplantation group (CT). Another 9 patients with 11 wounds transplanted with STSG admitted within the same time frame were enrolled as control group (C). All patients were followed up for longer than 2 years. Color, evenness, texture, contracture, sensation, and complications of transplanted skin were assessed using a modified Manchester Scar Scale (1-4 scores, the higher the score, the poorer the situation). The scar formation on skin donor sites was assessed by the Vancouver Scar Scale. Patients' degree of satisfaction and health status during the transplantation period were investigated in the form of questionnaire. The skin tissue structure of 4 patients was observed with histological method. The joint range of motion was assessed by the neutral position before and after operation and at follow-up. Data were processed with nonparametric test, chi-square test or t test.

RESULTS(1) The evenness, contracture, and texture of transplanted skin in CT group scored (1.6 ± 0.5), (1.8 ± 0.8), and (1.5 ± 0.8), respectively, which were significantly lower than those in C group [(2.0 ± 0.7), (2.2 ± 0.9), and (2.3 ± 0.7), with Z value respectively -2.058, -2.220, -2.323, P values all below 0.05]. Scores of color, sensation, and complications of transplanted skin in two groups were close to each other (with Z value respectively -0.628, -0.428, -2.520, P values all above 0.05). (2) Mild scar formation was observed in one of the skin donor sites in CT group. (3) Information as obtained from questionnaire showed no statistical difference between two groups in pinching, itching, and satisfaction degree (with χ(2) value respectively 0.187, 0.019, 2.628, P values all above 0.05). (4) Nerve fibers were seen in hand tissue 2 years after operation. ADM did not induce severe inflammatory responses in the site of grafting. (5) Eleven joints in CT group recovered or improved in function; while the other two joints required secondary surgery. Obvious contracture was observed in the two joints in C group.

CONCLUSIONSAllogeneic ADM combined with STSG transplantation prevents scar contracture and has obvious effect in improving function and appearance. There is no problem in regard to safety for its existence in either adult or children.

Adolescent ; Adult ; Burns ; surgery ; Child ; Child, Preschool ; Dermis ; transplantation ; Female ; Follow-Up Studies ; Humans ; Male ; Skin Transplantation ; methods ; Skin, Artificial ; Time ; Transplantation, Autologous ; Transplantation, Homologous ; Young Adult

OBJECTIVETo analyze the characteristics and the number of organs involved in acute graft-versus-host disease (aGVHD) among different donors of allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSClinical data were retrospectively analyzed in 289 patients received allo-HSCT in our hospital, between November 2007 and December 2008. Clinical features of the involved organs between different donors were compared.

RESULTSThe cumulative incidence of aGVHD was 57.4% (166/289), grades I-II and grades III-IV were 52.1% and 11.0%, respectively. Skin was involved in 116 (69.9%) of the total 166 cases, gut 97 (47.6%), and liver 25(15.1%). Organs involved in HLA-identical sibling transplantation and in haplo-identical ones were skin 19 (42.2%), gut 25 (55.6%), and liver 12 (26.7%), and 79 (80.2%), 54 (44.6%) and 13 (10.7%) respectively. More aGVHD involvements of skin were found in HLA haplo-identical HSCT than in HLA identical sibling HSCT (P = 0.000). The involvement of skin grade II was 8(17.8%) and 38.8% (P = 0.01), gut grade II was 22 (48.9%) and 36 (29.8%) (P = 0.028) in HLA identical sibling transplantation and in haploidentical HSCT, repectively. The incidences of aGVHD grades I to II and grade III to IV were 103 (62.0%) versus 12 (7.2%) in the single organ involved group, 37 (22.3%) versus 11 (6.6%) in the double organs involved group, and 0% versus 3 (1.8%) in the triple organs involved group.

CONCLUSIONThe percentage of aGVHD with skin involvement in haploidentical HSCT is significantly higher than that in HLA identical HSCT. There is a significant difference in mild skin or GI aGVHD between HLA matched and mismatched HSCT, but no difference in severe skin or GI aGVHD between the two groups. The percentage of severe aGVHD was higher if three organs were involved.

Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Retrospective Studies ; Siblings ; Tissue Donors ; Transplantation, Homologous

BACKGROUNDCyclin B1 (CLB1) is necessary for mitotic initiation in mammalian cells and plays important roles in cancer development. Therefore, a potential strategy in cancer therapy is to suppress the activity of CLB1 by delivering antisense constructs of CLB1 into tumor cells. In previous CLB1 studies, antisense constructs with a short half life were often used and these constructs might not persistently inhibit CLB1.

METHODSWe successfully created a recombinant plasmid encoding the full-length antisense cDNA of mouse cyclin B1 (AS-mCLB1) and transfected this construct to the murine Lewis lung carcinoma (LL/2) and CT-26 colon carcinoma (CT-26) cells. We isolated clones of LL/2 and CT-26 transfectants with stable expression of AS-mCLB1. Reverse transcriptional polymerase chain reaction (RT-PCR) and Western blot were applied to detect the expression of the mRNA and protein levels of CLB1. To further test the efficacy of this strategy in vivo, AS-mCLB1-expressing LL/2 and CT-26 transfectants were implanted into mice.

RESULTSWe found the expression of the mRNA and protein levels of CLB1 decrease in these transfectants. The inhibition of CLB1 caused prominent G1 arrest, abnormal morphology, retarded cell growth and an increase in apoptosis. In AS-mCLB1-expressing LL/2 and CT-26 transfectants implanted mice, tumorigenicity was effectively suppressed compared with the controls. In addition, the expression of AS-mCLB1 also significantly increases the survival duration of implanted animals.

CONCLUSIONAS-mCLB1 is likely to be useful in future cancer therapy, which may be associated with its ability to down-regulate the expression of CLB1 and then induce G1arrest and apoptosis in tumor cells.

Animals ; Apoptosis ; Cell Proliferation ; Cell Survival ; Cyclin B ; antagonists & inhibitors ; genetics ; Cyclin B1 ; DNA, Antisense ; pharmacology ; DNA, Complementary ; pharmacology ; G1 Phase ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Neoplasms, Experimental ; pathology ; therapy