Gene therapy has emerged as an efficient modality to treat human diseases.This method is based on the transfer of genetic material to tissues to induce a curative effect.Gene therapy vectors are molecular constructs used to facilitate the penetration of genomic sequences inside the cells.Viral vectots have however several limitations when administered directly to the patient.They may cause significant toxicity by activating innate immunity or by eliciting an adaptive immune response against viral proteins.In addition,targeting the vector to the desired site is an issue when given systemically.The use of cells as vehicles for gene therapy vectors has many advantages.The combination of cell-viro-gene therapy has been thought as a new and promising strategy for therapy of cancer.The targeting vector to cancer stem cells will become a new direction in the field of gene therapy.In this article,we will introduce progressions,limitations and future directions of gene therapy of liver cancer.

Objective To express recombinant HBHAΔC and HBHAΔN protein,and compare the HBHA series protein activity with each other.It will be provide a experimental basis for the research on clinical diagnostic of HBHA.Methods The HBHAΔC and HBHAΔN gene fragments were cloned and expressed by transforming E.coli BL-21.Test the protein heparin binding ability by CL-6B column.And then added protein to the BCG 7H9 culture medium,to observe the induced BCG aggregation.Results nHB-HA,rHBHA and HBHAΔN protein have heparin binding ability.Meanwhile nHBHA,rHBHA and HBHA Δ C protein have in-duced BCG aggregation effect.Conclusion The HBHAΔC and HBHAΔN protein were successfully obtained.It was proved that the HBHA C-terminal could be combined with heparin and the N-terminal involved could induce the aggregation of BCG.This results provide a basis for further study on molecular mechanism of TB infection and clinical application.

OBJECTIVETo investigate the feasibility and clinical results of applying poly-DL-lactic acid (PDLLA) biomembranes in cleft palate repair.

METHODS68 cleft palate patients were divided into study group and control group. The traditional surgical method was used to control group to close the soft cleft palate, and the PDLLA biomembrane was used to study group and implanted into the surgical gap between the periosteum and bone at the hard palate, and fixed with suture. The duration, blood loss at operation, post-operative complication, wound healing and recovery were recorded and compared to conventional cleft palate repair.

RESULTSOperations were successfully completed on all 34 patients. Wound healing of soft palate and uvula was uneventful with no incidence of fistula or dehiscence. The primary healing on tissue defect of hard palate occurred in 29 patients, secondary healing occurred in 3 patients, permanent fistula between the oral cavity and the nasal cavity occurred in only one patients, and 3 patients left over fistula on alveolar process. Compared to traditional cleft palate repair, blood loss and incidence of fistula on alveolar process were decreased; the average surgical time was 89.25 minutes and was not prolonged; and there was no significant increase in post-operative complication.

CONCLUSIONHard cleft palate repair with PDLLA biomembranes is safe, simple and practical with good clinical results and is beneficial to minimize the bad influences towards the development and growth for maxilla of cleft palate patients.

Absorbable Implants ; Adolescent ; Adult ; Child ; Child, Preschool ; Cleft Palate ; surgery ; Feasibility Studies ; Female ; Guided Tissue Regeneration ; methods ; Humans ; Lactic Acid ; therapeutic use ; Male ; Maxillofacial Development ; Palate, Hard ; surgery ; Polyesters ; Polymers ; therapeutic use ; Reconstructive Surgical Procedures

China ; epidemiology ; Circoviridae ; genetics ; isolation & purification ; Circoviridae Infections ; epidemiology ; virology ; DNA Virus Infections ; epidemiology ; transmission ; virology ; DNA Viruses ; genetics ; isolation & purification ; DNA, Viral ; blood ; Hepatitis Viruses ; genetics ; Humans ; Nucleic Acid Probes ; Polymerase Chain Reaction ; methods ; Sequence Homology, Nucleic Acid ; Transfusion Reaction

Objective To investigate whether paraptosis and autophagy have an effect on acute retinal ischemia-reperfusion injury (RIRI) in an experimental rat model that recapitulates features of acute hypertensive glaucoma and to explore the possible underlying mechanisms.Methods A total of 30 adult male Sprague-Dawley rats were randomly divided into RIRI group and control group.The acute RIRI model was induced with normal saline in the right eye of rats from the RIRI group by anterior chamber perfusion,while the rats in the control group left untreated.On day 1,day 3,day 7,day 28 after RIRI model establishment,the changes in morphology of retinal ganglion cells (RGCs) were observed by transmission electron microscopy (TEM),and the expression of microtubule-associated protein 1 light chain 3 (LC3) was measured by immumofluorescence methods.Results When compared with the control group,the number of cytoplasmic vacuoles predominantly derived from the progressive swelling of mitochondria and/or endoplasmic reticulum (ER) in RGCs were increased in the RIRI group from day 1 to day 28 by TEM.And ultra-structural analyses showed the double-or multiple-membrane autophagosomes were markedly accumulated in the cytoplasm of RGCs following acute RIRI.The average number of autophagic vacuoles in the cytoplasm of RGCs was 0.79 per 50 μm2 in the control group,and the average number of autophagosomes reached to a maximum on day 7 after acute RIRI at 2.29 per 50 μm2,which was statistically significant compared with the control group (P ＜ 0.05).Compared to the control group,LC3 expression in the cytoplasm of RGCs was up-regulated on day 1 after acute RIRI,which sustained throughout the experimental period.The percentage of LC3 positive cells in the retinal ganglion cell layer was 15.90％ in the control group,and the data was 46.95％ and 52.30％ on day 1 and day 28 after RIRI,respectively,both which were statistically significant compared with the normal control group (both P ＜ 0.05).Conclusion Both paraptosis and autophagy participate in death of RGCs after acute RIRI.Programmed cell death in different cells,either coexistence of multiple-cell death form or a single-cell death form,participates in the pathogenesis of acute RIRI.

BACKGROUNDIt is widely accepted that tumor necrosis factor-α (TNF-α) plays an important role in the pathogenesis of emphysema. This study aimed at investigating the protective effects of anti-TNF-α antibody, infliximab, in the development of emphysema induced by passive smoking in rats.

METHODSThirty-nine rats were randomly divided into a normal control group (group 1), an emphysema group (group 2), and an infliximab-intervention group (group 3). Rat models of emphysema were established by exposure to cigarette smoking daily for 74 days. After 1 month, the infliximab intervention group was treated with infliximab via subcutaneous injection. The levels of TNF-α, IL-8 and vascular endothelial growth factor (VEGF) in bronchoalveolar lavage fluid (BALF) were measured with enzyme linked immunosorbent assay (ELISA). The number and classification of cells in the BALF were measured. Lung tissue sections stained by hematoxylin and eosin (HE) were observed, and mean linear intercept (MLI) and mean alveolar numbers (MAN) were measured. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) methods were used to examine the percentage of positive cells and distribution of apoptotic cells.

RESULTSThe levels of TNF-α and IL-8 in BALF were higher in group 2 than in group 1 and group 3. The MLI was greater in group 2 than that in group 1 and group 3 while MAN was decreased. The concentration of VEGF in BALF of group 2 was significantly decreased as compared with group 1. The total cells and neutrophils number was significantly increased in group 2 as compared with group 1 and group 3, so was the percentage of neutrophils. The number of TUNEL positive cells in the alveolar septa was significantly increased in group 2 as compared with group 1 and group 3.

CONCLUSIONInfliximab protects against cigarette smoking-induced emphysema by reducing airway inflammation, attenuating alveolar septa cell apoptosis and improving pathological changes.

Animals ; Antibodies, Monoclonal ; therapeutic use ; Bronchoalveolar Lavage Fluid ; chemistry ; cytology ; Infliximab ; Interleukin-8 ; metabolism ; Male ; Pulmonary Alveoli ; cytology ; drug effects ; Pulmonary Emphysema ; chemically induced ; metabolism ; prevention & control ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tobacco Smoke Pollution ; adverse effects ; Tumor Necrosis Factor-alpha ; metabolism

Adult ; Asian Continental Ancestry Group ; genetics ; Female ; Humans ; Male ; Mutation ; Obesity ; genetics ; Receptor, Melanocortin, Type 4 ; genetics ; Young Adult

OBJECTIVETo study the effect of granulocyte colony-stimulating factor (G-CSF) on the proliferation and differentiation of bcr/abl(+)-CD34+ cells.

METHODSbcr/abl(+)-CD34+ cells were isolated from bone marrow of chronic myelocytic leukemia (CML) patients and were treated with 0, 10, 100, 1000 ng/ml of G-CSF for 48, 96, 144 hs. CD34 cells from normal bone marrow were used as controls. Cell proliferation was determined by trypan blue dye exclusion, cell-cycle and antigen differentiation were determined by flow cytometry and cell morphology was observed under light microscope.

RESULTSThe number of bcr/abl(+)-CD34+ cells was increased obviously in all groups. After cultured for 48 and 96 h, the number of bcr/abl(+)-CD34+ cells at G-CSF 10 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05) , the number of normal CD34 cells was increased only in the presence of G-CSF. After cultured for 48, 96 and 144 h, the cell number in G-CSF 100 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05, P < 0.01, P < 0.01, respectively). After cultured for 144 h, the cell percentages in G0/G1 phase for bcr/abl(+)-CD34+ cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that in G-CSF 0 ng/ml group (P < 0. 05), and that for normal CD34 cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that of G-CSF 0 ng/ml group after cultured for 48 and 96 h. The expressions of CD34 on bcr/abl(+)-CD34+ cells and normal CD34+ cells were decreased along with the culture duration, accompanied by the expression of CD33 and CD13 increased first and decreased later, which was not correlated with the concentration of G-CSF. Both bcr/abl(+)-CD34+ cells and normal CD34+ cells showed mature morphology along with proliferation and differentiation.

CONCLUSIONSG-CSF promotes proliferation of both bcr/abl(+)-CD34+ cells and normal CD34+ cells, but not necessary for the former, and the former differentiates more rapidly than the latter does, but both was independent of G-CSF.

Adult ; Aged ; Antigens, CD34 ; metabolism ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Female ; Fusion Proteins, bcr-abl ; metabolism ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; Male ; Monocytes ; cytology ; drug effects ; immunology ; Tumor Cells, Cultured

OBJECTIVETo investigate the factors influencing recurrence and metastasis following curative resection of pancreatic ductal adenocarcinoma and analyze the prognosis.

METHODSThe clinicopathological and follow-up data of 56 patients who underwent curative resection for pancreatic ductal adenocarcinoma between Jan. 1997 and Dec. 2006 in this hospital were analyzed retrospectively.

RESULTSThe recurrence rate after curative resection was 73.2% (41/56). The recurrence rate after operation at the time of 3 months, half year, 1 year and 2 years was 26.8% (15/56), 51.8% (29/56), 64.3% (36/56) and 69.6% (39/56), respectively. Hepatic metastasis and local recurrence accounted for 36.6% and 31.7% of the cases, respectively. The 3-year accumulated survival of this group was 22.7%. The symptom presenting time, back pain, preoperative level of CA19-9, tumor size, AJCC stage and T stage were correlated with metastasis/recurrence. Univariate analysis revealed that the preoperative level of CA19-9, T stage and the tumor size were prognostic factors. Cox regression analysis revealed that only tumor size was an independent prognostic factor.

CONCLUSIONThe metastasis or recurrence mostly occurs within 2 years after curative resection, and the liver is the most common site of metastasis. High recurrence rate is the major reason causing the failure of curative resection and short survival time after operation. The symptom presenting time, back pain, preoperative level of CA19-9, tumor size, AJCC stage and T stage are correlated with metastasis/recurrence. The tumor size is an independent prognostic factor.

Adult ; Aged ; CA-19-9 Antigen ; metabolism ; Carcinoma, Pancreatic Ductal ; immunology ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Pancreatectomy ; methods ; Pancreatic Neoplasms ; immunology ; pathology ; surgery ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate ; Tumor Burden

Objective To investigate the awareness rate,treatment rate and control rate of mineral and bone disorder in patients with moderate or advanced stage chronic kidney disease (CKD). Methods The awareness rate,treatment rate and control rate of mineral and bone disorder were evaluated based on a questionnaire and related laboratory examinations in 503 CKD stage 3 to 5 patients. Results The awareness rate of mineral and bone disorder in patients with moderate or advanced stage CKD was highest in hemodialysis patients,moderate in peritoneal dialysis patients and lowest in non-dialyzed patients (all P ＜0.01).The total scores of the questionnaire were lowest in non-dialyzed patients [6 (5,8)] and were significantly higher in peritoneal dialysis [11 (9,12)] and hemodialysis patients [13 (11,15)] (P＜0.01).The extent of awareness was negatively correlated with age (r=-0.11,P＜0.05),and positively correlated with educational background (r=0.226,P＜0.01),duration of CKD (r=0.597,P＜0.01) and duration of dialysis (r=0.366,P＜0.01).The source of knowledge was mainly from publicity and education made by medical staff,which accounted for 94.0％,79.5％ and 69.4％ respectively in nondialyzed,peritoneal dialysis and hemodialysis patients.The treatment rate was significantly higher in peritoneal dialysis (88.6％) and hemodialysis patients (96.9％) than that in non-dialyzed patients (58.2％) (all P＜0.01).According to K/DOQI guideline,the control rate of serum calcium,phosphorus,calcium and phosphorus product and parathyroid hormone (PTH) were much better in non-dialyzed patients as compared to dialyzed ones.The percentage of number of lab indicators meeting the standard was significantly higher in non-dialyzed patients as compared to dialyzed ones (P＜0.01).According to KDIGO guideline,the control rate of serum phosphorus was significantly lower in hemodialysis patients (23.6％) than that in peritoneal dialysis (36.9％) and non-dialyzed patients (46.7％) (P＜0.01). Conclusions In non-dialyzed patients with moderate or advanced stage CKD,the awareness rate and treatment rate of mineral and bone disorder are relatively low,and the control rate is relatively high.Whereas in dialyzed patients,the awareness rate and treatment rate are relatively high,and the control rate is relatively low.