Objective To construct recombinant prokaryotic plasmid DNA encoding mycobacterium tuberculosis (TB) heat shock protein 70 (HSP70) - Dnak gene obtained from pMT70, then purify its characterizes and the role of rHSP70 in tumor therapy. Methods Amplified by polymerase chain reaction (PCR) and with terminal modification, the Dnak gene was cloned into the vector with T7 promoter and was confirmed by restriction endonuclease digestion. So a new plasmid pMT70 - 3 was constructed and transformed in E. Coli. Strain BL21DE3. The engineering bacteria were induced by IPTG. Western-blot,natine-page and scanning were used to analyze the results. The human HSP70 were induced with peptides of lymphocytic leukemia cells of L1210 of mice in binding buffer which contain MgCl2 and ADP at 37 ℃ , then mice were given rHSP70 peptide - complex by SV injection twice at weekly intervals. Results The Dnak gene could be highly expressed in E. Coli as soluble protein. The expressing efficiency was 65% of the total cell protein and the soluble protein was 90% of the expressed protein. The rHSP70 - peptide complex could improve the immune protective function which had significant difference compared with controls (P

Professor WEI Pin-kang has engaged in prevention and treatment of gastrointestinal tumors through intrgrated traditional Chinese and Western medicine for more than forty years. According to the characteristics of the gastric cancer development and using the philosophy thinking of TCM comparative states, combined with the ancient theory of phlegm syndrome in TCM, Professor WEI Pin-kang put foward his original theory that is the concept of phlegm differentiation of gastric cancer theory, and built the theory of the stomach sputum pollution, which can renovate the body's environment, eradicating the cause and improving the quality of life for cancer patients.

Objective To observe the effects of iodine excess on thyroid morphology,the expression of thyroid peroxidase and sodium iodide symporter mRNA and to explore their mechanisms.Methods One-month SD rats were divided into three groups:control iodine(CI),high iodine Ⅰ(HI Ⅰ)and high iodineⅡ(HI Ⅱ)and were fed with water containing iodine in different concentrations by adding K103(5,5000,10 000μg/L)respectively.Rats were sacrificed after being fed for six months.The morphology of thyroid was investigated under light microscopy and electron microscopy,the serum thyroid hormones and ratio of TPO/β-actin and NIS/β-actin were measured by radio-immunoassay and RT-PCR method.Results The major changes were increased follicles with colloid accumulation in HI groups.The levels of serum thyroid hormones TT3 and TT4 were decreased gradually from CI[(75.68±13.99,1.45±0.49)nmol/L]to HI Ⅰ[(73.82±16.48,1.34±0.31)nmol/L]and HIⅡ groups[(70.65±11.43,1.15±0.39)nmol/L],but there were no significant differences among three groups(F=O.371,l.163,P＞0.05).The TPO and NIS mRNA expressions in HI Ⅰ(1.28±0.10,0.56±O.17)and HI Ⅱ(1.14±0.04,0.39±0.06)were significantly lower(F=30.863,62.62.675,P＜O.05)than those of control group(1.39±0.08,0.71±0.13).Conclusions Chronic iodine excess leads to definite histological changes in rat thyroid,and inhibits the expressions of TPO and NIS mRNA as well as thyroid hormone synthesis,which in turn acts as a protective mechanism against iodine excess.

Objective To explore the interaction characters of fluoride and aluminum by analyzing the changes of bone metabolism in mice. Methods Kunming mice were randomly divided into nine groups according to the factorial experiment, design of two factors and three levels. The animals in different groups were fed with various doses of fluoride(NaF, 0,50,150 mg/L) and/or aluminum(AlCl3, 0,200,600 mg/L) in drinking water for 24 weeks. Serum calcium, phosphor, magnesium, alkaline phosphatase, osteoealine, parathyroid, and urinary calcium and phosphor were tested. Results We found interaetians of fluoride and aluminum with serum calcium, osteoealine and urine calcium(F=17.370,4.399,9.448, P<0.01), but not with serum phosphor, magnesium, alkaline phosphatase, parathyroid or urinary phospbor(F=0.416,0.415,1.921,1.362, 1.630, P 0.05). The serum levels of calcium and osteoealine in high fluoride group were (1.13±0.27)mmol/L and (6.56±5.74)μg/L, respectively, which were lower than in the control group[ (1.82±0.37)mmol/L and (23.45±15.40)laeJL, respectively], but the levels were elevated to (1.76±0.36)mmol/L and (10.57±4.28)μg/L when high fluoride was combined with low aluminum, and further elevated to (2.10±0.51)mmol/L and (15.73±3.15)μg/L when high fluoride was combined with high aluminum. The urinary calcium level in low fluoride group [ (6.24±2.61)retool/retool Cr] was higher than that in the control group[ (3.12±2.04)retool/retool Cr], but it was decreased in low fluoride and aluminum groups[ (0.81±0.44), (1.23± 0.41)mmol/mmoi Cr, respectively]. On the other hand, the levels of serum ealeium and osteocaline in high aluminum group were (1.07±0.68)mmol/L and (7.21±5.22)μg/L, elevated to (1.47±0.18)mmol/L and (10.98±4.35) μg/L when low fluoride was combined wth high aluminum, and further elevated to (2.10±0.51)mmol/L and (15.73± 3.15)μg/L when high fluoride was.combined with high aluminum, respectively, and the combined effects showed the same trend of higher aluminum. Conclusions Aluminum antagonized fluoride-induced effects, whereas fluoride aggravated the effects caused by aluminum in this experimental conditions. The biomarkers of bone formation and mineralization were suppressed in the combined groups, so the combined effects could interfere with the course of bone turnover by inhabiting bone formation and mineralization, leading to the disorder of bone metabolism eventually.

Objective To investigate the combined effects of fluoride and aluminum intoxication on bones and their possible mechanisms.Methods Kunming mice were divided into nine groups according to the factorial experiment design.Different dose of fluoride(NaF,0,50,150 mg/L)and/or aluminum(AlCl3,0,200,600 mg/L)was administered to each group in drinking water.After 24 weeks,the degree of mottled teeth and the histomorphometric parameters,such as the bone trabecula and osteoid areas,the number of osteoblasts and osteoclasts,and pathologic changes in femur were observed.Results Aluminum could also caused mottled teeth(in degree 4).The mottled teeth in the combined groups were more serious than those in fluoride or aluminum alone group.The interaction between fluoride and aluminum existed in the changes of bone trabecula and osteoid areas(F=2.963,3.688,P＜0.05),and not existed in changes of the number of osteoblasts and osteoclasts(F=2.347,0.888,P＞0.05).In high fluoride group,the trabecula and osteoid areas were(50 675.47±22 916.34),(10 733.97 ±3015.55)μm2,but it increased to(75 988.64±13 797.21),(16 402.88±4605.83)μm2 when combined with high aluminum(P＜0.05),and the group of high fluoride +low aluminum increased to(69 277.16±19 837.51),(18 564.79±6362.47)μm2 (P＜0.05),so aluminum antagonized the effects induced by fluoride;the area of bone trabecula of group of high aluminum was(60 718.43 ±17 574.37)μm2,but it increased[(75 988.64±13 797.21),(82 474.94±15 466.66)μm2]when combined with high or low fluoride(P＜0.05),and the combined effects showed a similarity to those in high aluminum group.The prominent osteoporosis with increased osteoid and cartilage tissues,and decreased amount of bony matrix and minerals were the main histopathological changes in the bone.Conclusions Both high aluminum and fluoride intoxication can result in mottled teeth,their combined effects are more serious than the individual effect.The prominent injury of combined fluoride and aluminum intoxication is osteomalacia and osteoporosis.

Specific primers 1CaA/1CaB for full cry1C gene in Bacillus thuringiensis subsp.colmeri strain 15A3 were designed. The 4.0kb PCR product included the whole ORF and regulation region of cry1C gene. This PCR product was linked with shuttle vector pHT315 by two cloning steps. The recombined plasmid pHT-1C was electroporated into Bacillus cereus 9509, a kind of bacteria that beneficial to crops. The transformant could produce bipyramidal-shaped parasporal inclusions. The 60kD protein band was detected by SDS-PAGE. The bioassay result showed that the cry1C gene transformant of Bc 9509 had insecticidal activity to Spodoptera exigua.

In this study, the buffering capacity of amphiphilic pH-sensitivity copolymer poly(2-ethyl-2-oxazoline)-cholesteryl methyl carbonate (PEOZ-CHMC) was evaluated. The ammonium sulfate gradient method was used to prepare doxorubicin hydrochloride (DOX x HCl)-loaded liposomes (DOX-L), and then the post-insertion method was used to prepare PEOZ-CHMC and polyethylene glycol-distearoyl phosphatidyl ethanolamine (PEG-DSPE) modified DOX x HCl-loaded liposomes (PEOZ-DOX-L and PEG-DOX-L). The physico-chemical properties, in vitro drugs release behavior, cellular toxicity and intracellular delivery of liposomes were evaluated, separately. The results showed that PEOZ-CHMC has a satisfactory buffering capacity. The sephadex G-50 column centrifugation method and dynamic light scattering were used to determine the encapsulation efficiency (EE) and particle size of liposomes. The EE and particle size of DOX-L were (97.3 ± 1.4) % and 120 nm, respectively, and the addition of PEOZ-CHMC or PEG-DSPE had no influence on EE and particle size. The zeta potentials of three kinds of liposomes were negative. The release behavior of various DOX liposomes in vitro was investigated by dialysis method. In phosphate buffer solution (PBS) at pH 7.4, DOX x HCl was released from PEOZ-DOX-L in a sustained manner. While in PBS at pH 5.0, the release rate of DOX x HCl from PEOZ-DOX-L increased significantly, which suggested DOX x HCl was released from PEOZ-DOX-L in a pH-dependent manner. The intracellular delivery of liposomes was investigated by confocal laser scanning microscopy (CLSM). The CLSM images indicated that PEOZ-DOX-L showed efficient intracellular trafficking including endosomal escape and release DOX x HCl into nucleus, as well as the DOX-L and PEG-DOX-L had no this effect. The cytotoxicity of liposomes against MCF-7 cells was detected by using MTT assay. The results showed that antiproliferative effects of PEOZ-DOX-L enhanced with pH value decreased, whereas DOX-L and PEG-DOX-L did not have any significant difference in inhibitions at different pH conditions. Therefore, the problems of the inhibition of cellular uptake of liposomes and the failed endosomal escape of pH-sensitive liposomes by PEG chain can be overcome by the pH-sensitive liposomes constructed by PEOZ-CHMC.

Self-cloning is a favorable technique in modification of industrial microorganisms especially food microorganisms because of its bio-safety. A survey and outlook of research in breeding of self-cloning industrial brewing yeast were introduced in this study.

In this study, the buffering capacity of amphiphilic pH-sensitivity copolymer poly(2-ethyl-2-oxazoline)-cholesteryl methyl carbonate (PEOZ-CHMC) was evaluated. The ammonium sulfate gradient method was used to prepare doxorubicin hydrochloride (DOX x HCl)-loaded liposomes (DOX-L), and then the post-insertion method was used to prepare PEOZ-CHMC and polyethylene glycol-distearoyl phosphatidyl ethanolamine (PEG-DSPE) modified DOX x HCl-loaded liposomes (PEOZ-DOX-L and PEG-DOX-L). The physico-chemical properties, in vitro drugs release behavior, cellular toxicity and intracellular delivery of liposomes were evaluated, separately. The results showed that PEOZ-CHMC has a satisfactory buffering capacity. The sephadex G-50 column centrifugation method and dynamic light scattering were used to determine the encapsulation efficiency (EE) and particle size of liposomes. The EE and particle size of DOX-L were (97.3 ± 1.4) % and 120 nm, respectively, and the addition of PEOZ-CHMC or PEG-DSPE had no influence on EE and particle size. The zeta potentials of three kinds of liposomes were negative. The release behavior of various DOX liposomes in vitro was investigated by dialysis method. In phosphate buffer solution (PBS) at pH 7.4, DOX x HCl was released from PEOZ-DOX-L in a sustained manner. While in PBS at pH 5.0, the release rate of DOX x HCl from PEOZ-DOX-L increased significantly, which suggested DOX x HCl was released from PEOZ-DOX-L in a pH-dependent manner. The intracellular delivery of liposomes was investigated by confocal laser scanning microscopy (CLSM). The CLSM images indicated that PEOZ-DOX-L showed efficient intracellular trafficking including endosomal escape and release DOX x HCl into nucleus, as well as the DOX-L and PEG-DOX-L had no this effect. The cytotoxicity of liposomes against MCF-7 cells was detected by using MTT assay. The results showed that antiproliferative effects of PEOZ-DOX-L enhanced with pH value decreased, whereas DOX-L and PEG-DOX-L did not have any significant difference in inhibitions at different pH conditions. Therefore, the problems of the inhibition of cellular uptake of liposomes and the failed endosomal escape of pH-sensitive liposomes by PEG chain can be overcome by the pH-sensitive liposomes constructed by PEOZ-CHMC.
Cell Nucleus ; Doxorubicin ; analogs & derivatives ; chemistry ; Endosomes ; Formates ; chemistry ; Humans ; Liposomes ; chemistry ; MCF-7 Cells ; Microscopy, Confocal ; Particle Size ; Phosphatidylethanolamines ; Polyamines ; chemistry ; Polyethylene Glycols ; chemistry

AIM: To measure the anatomical character of nasolacrimal dust by spiral CT 3D reconstruction in children.METHODS: The length of nasolacrimal dust and the angles between nasolacrimal dust and vertical plane,horizontal plane and coronal plane in 27 children(54 eyes) and 15 adults(30 eyes) were surveyed using spiral CT 3D reconstruction. While the length of nasolacrimal dust in 7 dead children(14 eyes) body were surveyed as comparison by anatomical method. The RESULTS:were analyzed by software SPSS 13.0 statistically. RESULTS: The length of nasolacrimal dust was 10.06±0.29mm in children and 11.51±1.54mm in adults by spiral CT 3D reconstruction,as 9.95±0.31mm in dead children body by anatomical method,with significant statistical difference between that of children and adults. The angles between nasolacrimal dust and vertical plane,horizontal plane and coronal plane in children were 7.96°±1.62°,73.24°±6.75°,and 12.31°±2.03° respectively,while the corresponding angels in adults were 8.08°±0.63°,72.69°±3.85° and 12.09°±1.21°. The difference between them had no statistical meaning. CONCLUSION: The anatomical data of children nasolacrimal dust obtained from spiral CT 3D reconstruction have important guidance to the therapy of nasolacrimal dust diseases in children.