Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.

In this article, the authors firstly summarized the number of applications submitted to and projects supported by the National Natural Science Foundation of China (NSFC) in the field of traditional Chinese medicine research in 2010. Then they described the district distribution, research direction layout and allotment of the approved projects in the three primary disciplines (traditional Chinese medicine, Chinese materia medica and integrated traditional Chinese and Western medicine) and their 43 subdisciplines. The targeting suggestions for improvement were given respectively by concluding the reason of disapproved projects from the point of view of applicants and supporting institution, and by stating the common problems existing in the review process from the perspectives of fund managers and evaluation experts. Lastly, the major funding fields in the near future were predicted in the hope of providing guidance for applicants.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Scalp ; Sleep Initiation and Maintenance Disorders ; therapy ; Treatment Outcome ; Young Adult

Objective To investigate the tissue stimulation of copper ions to the smooth muscle of murine terminal rectum.Methods In this study copper needle used for electrochemical therapeutic equipment was punctured into submucosal smooth muscle tissue of terminal rectum above the dentate line in 65 male rats.Rats were then divided into 5 groups (10 rats in each group) at random and sacrificed at 1, 3, 4, 6, 8 weeks after respectively, samples were sent for observation of macroscopic and microscopic tissue reaction.Results Rats had no abnormal histological change in the puncture points grossly. Undor microscope, mild edema was detected in the submucosal layer in 1 week group. Remarkable infiltration of inflammatory cells and lymphocytes subsided 3 weeks after, and disappeared five weeks after.Smooth muscle was normal microscopically in all groups.In contrast, platinum and steel needles caused infiltration of plasma cells and neutrophils, ulcers and small abscess formation in around puncture points.Conclusion Inflammatory reaction was induced after puncturing of copper needle into the tissues and the tissue reaction disappeared after 5 Ws.In contrast with platinum and steel needles, copper needle was non-traumatic to the smooth muscle tissue.

AIM: To discuss the relationship between prostate specific membrane antigen(PSMA) and prostate cancer and to seek a target for diagnosis and therapy of prostate cancer.METHODS: A pair of primers was designed according to the published PSMA mRNA sequence.Total RNA was extracted from prostate cancer tissues and was reversely transcribed into cDNA,which was used as a template for PCR to amplify the PSMA gene.The recombinant was sequenced and the result was analyzed by BLAST.The PSMA5 gene specific primers were designed to identify its expression in different cells and prostate tissues.RESULTS: A new alternatively spliced variant of PSMA named PSMA5 was discovered when sequencing the recombinant.PSMA5 showed well pathological tissue-specificity,and its expression rate in prostate cancer,benign prostatic hyperplasia of prostate,and normal prostate tissue were 92.6%,78.8% and 10.0%,respectively.It expressed specifically in Pca cell line LNCaP,not in cell lines of PC3,bladder carcinoma,renal carcinoma,or hepatoma.CONCLUSION: A new alternative spliced variant of PSMA named PSMA5 was discovered,which was well correlated with prostate cancer and benign prostatic hyperplasia.This finding may give a new clue to the evolution of prostate cancer and may provide a target for the diagnosis and therapy of prostate cancer.

AIM: To find out the gene structure and diversity of protate specific membrane antigen(PSMA) alternative spliced variants, and probe into the pathogenesis of prostate cancer.METHODS: 5'-RACE and 3'-RACE methods were used to amplify the 5' and 3'end of alternative spliced variant and then those viariants were sequenced for analyzing the gene stucture and diversity of PSMA alternative spliced variants of prostate cancer tissues.RESULTS: Four new alternative spliced variants of PSMA were discovered from prostate cancer tissues.Compared with reported PSMA alternative spliced variants,different insertions and deletions existed in different sites of those new variants.CONCLUSION: The discovery of the new variants confirms the diversity of PSMA spliced variants and provides the clues for seeking the target of diagnosis and therapy of prostate cancer.

Objective To analyze the clinical characteristics of adult patients with hemophagocytic lymphohistiocytosis (HLH) receiving haploidentical donor hematopoietic stem cell transplantation (HID HSCT).Method We retrospectively reviewed 20 adult patients with HLH from August 2009 to August 2014.The clinical features and outcome were analyzed.Results Conditioning regimens consisted of total body irradiation/etoposide/cyclophosphamide (TBI/VP-16/CTX) and busulfan (Bu)/VP-16/CTX in HLH with anti-thymocyte globulin (ATG) 8 mg/kg.The stem cells were mobilized from donors' peripheral blood.Median time to white blood cell engraftment was 13 (9-27) days.Median time to platelet engraftment was 14 (10-28) days.Mixed chimerism after transplantation developed in 4 patients and no patient presented graft failure.Eight patients developed grade Ⅱ to Ⅲ acute graft-versus-host disease (GVHD),while as chronic GVHD occurred in 9 patients.Among 12 patients with EB virus (EBV) reactivation,2 patients developed post-transplant lymphoproliferative disorder (PTLD),7 were suspected as PTLD and 3 were considered as relapse of primary disease.With a median follow-up of 20 months (range:0.5-108 months) after transplantation,the estimated 2-year overall survival (OS) rate was (60.0 ± 11.0) ％ in all patients.During the follow-up,12 patients survived,8 died including 5 within 100 days after HSCT.Among 5 non-remission patients before HSCT,4 patients died within 100 days after HCT.Conclusions HID HSCT is an effective treatment for adult patients with HLH to achieve remission and long-term survival.High proportion of mixed chimerism has been seen at early stage after transplantation.EBV reactivation and early transplant-related mortality are common.

BACKGROUND:Many experiments and clinical studies have reported that venous blood, absorbable colagen sponge and rich fibrin clot, without the use of bone graft material, can al promote new bone formation; therefore, whether only transplanting concentrated growth factor in the maxilary sinus lift could effectively promote bone regeneration? OBJECTIVE: To conduct the maxilary sinus lift with concentrated growth factor as the graft material, and to observe the peri-implant bone level change. METHODS: Totaly 26 patients were involved, including 14 males and 12 females, aged 35-73 years. Maxilary sinus lift was conducted and autologous concentrated growth factor was taken as the graft material. Astra Tech implants were implanted simultaneously. Patients were divided into 6-12 months, 13-18 months and > 18 months groups according to the folow-up time. Patients were divided into≤ 5 mm, 5-7 mm, and≥ 7 mm groups according to the preoperative alveolar bone height. Patients were divided into≤ 2 mm, 2.0-3.0 mm, and≥3 mm groups according to the postoperative maxilary sinus lift height. The survival rate of implants and marginal bone level changes were observed during the folow-up. RESULTS AND CONCLUSION: Totaly 44 implants were implanted, of which 43 implants were considered as successful repair during the 6-18 months of postoperative folow-up. The survival rate of implants was 98%. There were no significant differences in the changes of mesial, distal and middle marginal bone levels between different folow-up time groups. The maxilary sinus lift with transplantation of concentrated growth factor resulted in a good osseointegration within 6-18 months post-operation. In addition, no significant differences were found in the changes of mesial, distal and middle marginal bone levels between different residual alveolar bone height groups, as wel as between different maxilary sinus lift height groups. It showed that the residual alveolar bone height and maxilary sinus lifting height had no significant effect on the peri-implant bone formation. Briefly, these findings demonstrate that the maxilary sinus lift with concentrated growth factor transplantation can result in a high survival rate of implants that are simultaneous implanted, and the alveolar bone around the implant is stable. But the long-term effect needs further observation.

Objective To determine the effect of reducing the value of mA or kV on the image quality and the radiation dose of the patients undergoing low-dose spiral CT for orthodontics.Methods Thirty patients were divided into three groups,each group has 10 patients.They were group 1(80 kV and 200 mA),group 2 (120 kV and 80 mA),group 3(120 kV and 200 mA)The volume CT dose index(CTDI)was recorded and the average dose-length produce(DLP)was calculated in three groups,respectively.Image quality of three groups were compared and scored by two radiologists,and the results were statistically analysed.Results The CTDI among three groups in diagnostic image quality.Conclusions Low-dose spiral CT for orthodontics,especially the low-kV scan,may decrease the radiation exposure and guarantee the image quality.

[Objective] To detect the expression of CD137L mRNA and protein in colorectal cancer tissues and discuss its possible clinical significance.[Methods] To detect the expression of CD137L mRNA in colorectal cancer tissues and adjacent noncancerous tissues by real-time RT-PCR and immunohistochemistry and to analyze the relationship between CD137L expression and clinical features.[Results] The mean level of CD137L mRNA in colorectal cancer tissues is significantly higher than that of adjacent noncancerous tissues (0.035 ± 0.013 vs.0.008 ± 0.005,P ＜ 0.05 ).CD137L mRNA expression in colorectal cancer show no significant differences among different age,sex,tumor location,Dukes classification,histological differentiation,and serum levels of CEA.We selected 8 colorectal cancer tissues and 3 adjacent tissues randomly to analyze the expression of CD137L by immunohistochemistry.CD137L was expressed on tumor cells in colorectal cancer tissues.The expression of CD137L was observed only in colorectal cancer tissues (6/8),but not in adjacent noncancerous tissues (0/3).[Conclusions] The expression of CD137L Mrna in colorectal cancer tissues is higher than that of adjacent noncancerous tissues and CD137 is expressed on tumor cell surface,it might be of biological importance for the progression of colorectal cancer.