Objective: To observe the clinical efficacy of acupoint injection with salmon calcitonin for back pain in elderly patients with primary osteoporosis. Methods: A total of 76 patients were selected and randomly divided into two groups by the random number table method, with 39 cases in the treatment group and 37 cases in the control group, respectively. Patients in both groups received routine anti-osteoporosis treatment. Patients in the treatment group received additional acupoint injection with salmon calcitonin at bilateral Pishu (BL 20) and Shenshu (BL 23), while patients in the control group received additional intramuscular injection with salmon calcitonin. The treatments for both groups were given once a day and lasted for 4 weeks. Visual analog scale (VAS) and Chinese Oswestry disability index (CODI) scores were observed before treatment, after 2 weeks and 4 weeks of treatment, and the use of analgesics during the treatment were recorded. Results: After treatment, the clinical efficacy in the treatment group was more significant than that in the control group (P<0.05). After 2 weeks and 4 weeks of treatment, the VAS scores in both groups showed significant intra-group and between-group differences (all P<0.05), and the CODI scores in both groups showed significant intra-group differences (all P<0.05). After 2 weeks of treatment, the CODI score showed no significant between-group difference (P>0.05). After 4 weeks of treatment, the improvement of CODI score in the treatment group was more significant than that in the control group (P<0.05). During the treatment, 2 cases in the treatment group took analgesics versus 8 cases in the control group, and the result showed a significant between-group difference (P<0.05). Conclusion: For back pain in elderly patients with primary osteoporosis, based on the routine treatment of oral medication, the clinical efficacy of acupoint injection with salmon calcitonin at bilateral Pishu (BL 20) and Shenshu (BL 23) is more significant than that of intramuscular injection. Acupoint injection treatment can improve patients' conditions and reduce the use of analgesics.

Adult ; Electromyography ; Gasoline ; poisoning ; Humans ; Male ; Poisoning ; physiopathology ; psychology ; Young Adult

Objective To investigate the effects of CD4+T cell depletion in BALB/c mice on the immunogenicity and virulence of replication-competent recombinant vaccinia virus. Methods Twelve BALB/c mice were inoculated with recombinant Tiantan vaccinia ( rTV, n=8 ) or vaccinia virus Tiantan strain ( VTT, n=4) by tail scarification after the depletion of CD4+T cells with anti-CD4 monoclonal anti-body( McAb) injected intraperitoneally for three days before virus inoculation.A control group without anti-body treatment was set up accordingly.Several parameters including the body weight, pocks on tail, viral shedding and the percentage of CD4+T cells were monitored.In the fourth week after virus infection, ovaries were collected from mice and viral loads in the tissue were titrated by plaque forming assay on chick embryo fibroblast ( CEF) cells.The specific cellular immune responses against vaccinia virus and HIV induced by inoculation of VTT and rTV respectively were detected by intracellular cytokine staining ( ICS) assay.ELISA was used to detect antibodies against vaccinia virus and HIV-1 gp120.Results All mice with or without CD4 McAb treatment showed typical poxvirus pocks on tails after inoculation of vaccinia viruses, but none of them developed secondary or satellite lesions.It took a longer time for CD4+T cell-depleted mice to heal from lesions, to regain body weights and to release viruses than the mice in control group.No vaccinia virus was detected in the ovaries of CD4+T cell-depleted mice or mice in control group.The mean absorbance( A) values for the detection of HIV-specific and vaccinia virus-specific antibodies in CD4+T cell-depleted mice with ELISA were respectively 0.119 and 0.168, which were significantly lower than those in mice of control group.The titers of neutralizing antibodies against vaccinia virus in McAb/rTV treated mice (1 ∶321) were lower than those in rTV treated mice (1 ∶1286) (P<0.05).The percentages of CD4+T cells secreting IFN-γ(0.654%) in McAb/rTV treated mice were significantly lower than those in rTV treated mice in the fourth week after immunization (P <0.0004).No significant differences with the vaccinia virus-specificCD8+ T cell responses were observed among mice with or without CD4+T cells depletion.Conclusion Thereplication and dissemination of replication-competent recombinant vaccinia virus could be effectively controlledin the mice with CD4+ T cell-depletion.The depletion of CD4+ T cells significantly reduced the humoraland CD4+ T cell responses, but had no effect on CD8+ T cell responses.

Objective To analyze the antibody responses in guinea pigs vaccinated with recombi-nant vaccinia virus( rTT) strains expressing transmitted/founder ( T/F) HIV-1 membrane proteins in combi-nation with gp140 protein.Methods Guinea pigs were primed with rTT strains and boosted twice with gp140 protein in every four weeks.Serum samples were collected from guinea pigs before immunization and in 2, 6 and 10 weeks after the last immunization for the detection of HIV-1-specific binding antibodies, neu-tralizingantibodiesandtherelativeavidityofantibodies.Results (1)Thebindingantibodiesspecificto HIV-1 B′/C, B, AE subtypes were efficiently induced by the immunization of rTT-B, rTT-C and rTT-CON vaccinia strains in combination with gp140 protein.The antibody titers ranged from 111 430 to 1 024 000. More antibodies against HIV-1 B′/C and AE subtypes were induced in guinea pigs by the immunization of rTT-C and rTT-CON strains in combination with gp140 protein than those by using rTT-B strain prime-protein boost strategy (P<0.05).No significant differences with the titers of HIV-1 B subtype specific antibody were observed among the guinea pigs immunized with the three strategies.( 2 ) High titers of SF162 and ZM109 neutralizing antibodies were induced in guinea pigs immunized with rTT-B, rTT-C and rTT-CON vac-cinia strains in combination with gp140 protein, ranging from 83.76 to 649.30.No significant differences were found among the three groups.(3) The HIV-1 V1V2-gp70 specific antibodies associated with protec-tive immunity were induced by immunization of the three virus prime-protein boost strategies.No significant differences were observed among them.(4) Antibodies induced in guinea pigs by immunization of the three strategies showed strong affinity to membrane proteins of HIV-1 B′/C, B, AE subtype strains.No significant differences were found among the three immunization strategies.Conclusion A strong humoral immune re-sponse was induced in guinea pigs primed with recombinant vaccinia virus strains expressing T/F virus HIV-1 membrane proteins and boosted with gp140 protein.

Objective To assess the prevalence and binocular dysfunctional risk factors associated with asthenopia among adult myopes.Methods The study population included 800 adult myopes,a cross-sectional visual parameters that characterize the accommodative:accommodation amplitude (AA),accommodative facility,and accommodative response (fused crossed cylinder-FCC) and binocular function (near and distant horizontal and vertical associated phorias,near and distance negative and positive fusional vergence,near point of convergence,negative and positive relative accommodation (NRA/PRA),stimulus AC/A ratio and stereoacuity) were evaluated when these subjects wore adequate spectacle correction.Results Asthenopia was reported in 24.2％ (194/800) of myopes.The incidence of asthenopia in female (27.8％,128/460) was more than that in male (19.4％,66/340),and there was significant difference (P =0.006).In univariate analysis,the monocular AA,binocular AA,NRA and PRA were significantly associated with asthenopia (P =0.000).In multivariate analysis,low NRA (≤1.25 D),low NRA (≤1.50 D) were significant risk factors for asthenopia (P =0.000,OR =7.644 ;95％ CI 2.913-17.580;P =0.000,OR =5.303;95％ CI 2.822-16.205).Conclusion Preventive measures directed against the binocular dysfunctional risks factors associated with asthenopia may help reduce the prevalence and provide a positive impact on asthenopia.

The positive ratio is 74 percent using the plate anaerobe culturing device made by author, higher than using common anaerobic box and anaerobic jar.

miR-200c has been shown to regulate the epithelial-mesenchymal transition (EMT) by inhibiting ZEB1 and ZEB2 expression in breast cancer cells. This study further examined the role of miR-200c in the invasion and metastasis of breast cancer that goes beyond the regulation on ZEB1 and ZEB2 expression. In this study, the bioinformatics software (miRanda) was used to predict the target gene of miR-200c and Renilla luciferase assay to verify the result. The metastatic breast cancer cells MDA-MB-231 were cultured and transfected with the miR-200c mimic or inhibitor. The expressions of miR-200c and HMGB1 were detected by RT-PCR and Western blotting, respectively. Transwell assay and wound healing assay were employed to examine the invasive and migrating ability of transfected cells. Target prediction and Renilla luciferase analysis revealed that HMGB1 was a putative target gene of miR-200c. After transfection of MDA-MB-231 cells with the miR-200c mimic or inhibitor, the expression of miR-200c was significantly increased or decreased when compared with cells transfected with the miR-200c mimic NC or inhibitor NC. Moreover, the expression of HMGB1 was reversely correlated with that of miR-200c in transfected cells. Tranwell assay showed that the number of invasive cells was significantly reduced in miR-200c mimic group when compared with miR-200c inhibitor group. It was also found that the migrating ability of cells transfected with miR-200c mimics was much lower than that of cells transfected with miR-200c inhibitors. It was suggested that miR-200c can suppress the invasion and migration of breast cancer cells by regulating the expression of HMGB1. miR-200c and HMGB1 may become useful biomarkers for progression of breast cancer and targets of gene therapy.

To reduce the serum clearance of interferon alpha2b, a chimeric gene encoding an human serum albumin(HSA)--human interferon alpha2b(IFNalpha2b) fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN alpha and anti-HSA antibody, was purified from the culture of the recombinant yeast by ultrafiltration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNa2b moiety exhibits antiviral activity similar to that of recombinant human IFNa2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90 microg/kg injection intravenously or subcutaneously. The elimination phase half-life of the fusion protein was 101h after intravenous injection and 68.2h after subcutaneous injection. Its Subcutaneous bioavailability was 67.9%. The enhanced pharmacokinetics of interferon a2b fused to human serum albumin suggest its promissing application in clinic medicine.
Animals ; Bioreactors ; microbiology ; Fermentation ; Humans ; Interferon-alpha ; biosynthesis ; genetics ; Macaca fascicularis ; Pichia ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacokinetics ; Recombinant Proteins ; Serum Albumin ; biosynthesis ; genetics

OBJECTIVETo compare the effectiveness of using multiple ligation probe amplification (MLPA) and denaturing high-performance liquid chromatography (DHPLC) in screening the exon deletions and duplications of the DMD gene.

METHODSMLPA technique was applied to detect exon deletions and duplications previously confirmed by denaturing high-performance liquid chromatography (DHPLC).

RESULTSFrom October 2004 to October 2005, 22 unrelated DMD probands and their possible female relatives with clinical diagnosis with dystrophinopathy at our hospital entered this study. Both DHPLC and MPLA detected DMD gene depletion in 11 probands and DMD duplications in 3 probands. MLPA detected deletions and duplications in 2 probands, which were not detected by DHPLC. MLPA also successfully identified the carriage status of the potential female carriers of the probands.

CONCLUSIONCompared with DHPLC and traditional PCR techniques, MLPA is a superior tool to analyze the deletions and duplications in affected males as well as in the identification of the carriage status of potential females carriers.

Chromatography, High Pressure Liquid ; Female ; Gene Deletion ; Gene Duplication ; Genetic Predisposition to Disease ; Humans ; Male ; Muscular Dystrophy, Duchenne ; genetics ; Mutation ; Nucleic Acid Amplification Techniques ; methods

OBJECTIVETo explore polarization of T lymphocyte and its relationship with apoptosis of marrow cells in patients with myelodysplastic syndromes (MDS).

METHODSMeasurements of Th1, Th2, Tc1, Tc2 subsets in bone marrows from 34 patients with MDS and 13 normal controls were performed by flow cytometry. INF-gamma and TNF-alpha in marrow serum were determined by ELISA (Enzyme-linked immunosorbent assay). Apoptosis index of marrow cells was detected by TUNEL (TdT-mediated dUTP nick end labeling). Correlations between Th1, Th2, Tc1, Tc2 subsets and INF-gamma, TNF-alpha levels as well as apoptosis index were analyzed, and relationship between TNF-alpha, INF-gamma levels and apoptosis index was also investigated.

RESULTS(1) The percentage of Th1 cells [(10.1 +/- 1.6)%], Tc1 cells [(24.0 +/- 3.6)%] and Tc1/Tc2 ratio (50.0 +/- 11.1) was significantly increased in patients with MDS than in normal controls [(4.0 +/- 0.5)%, (5.8 +/- 0.6)% and 13.4 +/- 2.7, respectively]. Levels of INF-gamma [(58.6 +/- 21.7) microg/L] and TNF-alpha [(15.7 +/- 3.8) microg/L] in marrow serum of MDS patients was markedly elevated compared to normal controls [0 and (0.3 +/- 0.2) microg/L, respectively]. An increased apoptosis index of nucleated cells was observed in MDS patients [(7.8 +/- 1.5)%] as compared to controls [(2.1 +/- 0.3)%, P < 0.05]. The Th1 cell percentage showed a positive correlation with the levels of INF-gamma and TNF-alpha (r = 0.38, P < 0.05 and r = 0.39, P < 0.05, respectively), and with apoptotic index of nucleated marrow cells in MDS patients (r = 0.33, P < 0.05). Furthermore, a positive correlation was observed between INF-gamma, TNF-alpha levels and apoptotic index of marrow cells (r = 0.74, P < 0.01 and r = 0.73, P < 0.01, respectively). (2) Th1, Tc1 cells and Tc1/Tc2 ratio in MDS-RCMD patients was markedly elevated (P < 0.01) but did not in RCMD-RS, RAEB-1 and RAEB-2 patients as compared to normal controls. (3) An elevation in the percentages of Th1, Tcl and Tc1/ Tc2 ratio was detected in patients with IPSS lower-risk but did not in higher-risk group as compared to controls. (4) Increased Th1 and Tc1 percentages and Th1/Th2 and Tc1/Tc2 ratios were observed in RCMD patients with normal karyotype, but did not in those with abnormal karyotype. Conclusions Th1/Th2 and Tc1/Tc2 in bone marrow of MDS patients were unbalanced, polarizing to type I reaction especially in patients with RCMD subtype, IPSS lower-risk and normal karyotype. The increased Th1 cells in bone marrow may account for the increased apoptosis of nucleated marrow cells in MDS, through proapoptotic cytokines such as INF-gamma and TNF-alpha.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apoptosis ; immunology ; Bone Marrow ; immunology ; metabolism ; pathology ; Female ; Hematopoietic System ; immunology ; Humans ; Interferon-gamma ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; T-Lymphocytes ; immunology ; Tumor Necrosis Factor-alpha ; metabolism