Objective To investigate the clinical features of the childhood chronic myelogenous leukemia (CCML), including the pathogenesis, incidence, clinical characteristics, diagnostic criterion, prognostic significance and the treatment strategies, etc. Method The data of 148 cases of CCML were comprehensively reviewed and analyzed, and international and domestic literature in the last two decades was reviewed. Results The CCML was found to be rare with unknown etiology, and was an acquired malignant disease of clonal proliferation of hematopoietic stem cells in children. The disease included two clinical types: adult CCML and juvenile CCML. 72.3% of CCML patients were diagnosed as the adult CCML. The clinical feature of CCML consisted of fatigue, low fever, anemia, hepatomegaly, splenomegaly, and lymphadenopathy. The laboratory findings of a typical CCML patient comprised of peripheral blood leukocytosis, basophilia and eosinophilia, myeloid differentiation in different stages, and increased megakaryocytes. The immunohistochemical features of the CCML consisted of highly positive MPO and CD68, significant lowering of neutrophil alkaline phosphatase (NAP), positive for Philadelphia chromosome (Ph) or chimeric BCR/ABL gene, etc. But in most cases of juvenile CCML, the Philadelphia chromosome could not be detected. The Gleevec therapy and hematopoietic stem cell transplantation (HSCT) might give better treatment result for CCML than the traditional therapy. Conclusions CCML has its characteristic clinical feature. The key of good therapeutic result is early diagnosis and treatment. The optimal therapy for CCML is Gleevec regime and HSCT.

Objective To construct short-hairpin RNA (shRNA) eukaryotic express vectors targeting of insulin like growth factor-1 receptor (IGF1R) gene, and to explore the changes of adhesion, invasion and FAK protein expression of MHCC-97 H hepatocellular carcinoma cells with RNA interference. Methods The shRNA oligonucleotide fragments were designed and synthesized based on the sequence of IGF1R mRNA. Double strands were formed after annealing and inserted into pGCsi-U6-Neo-GFP vector. The recombinant was transformed into Stable 3, then plasmids were extracted and identified by restriction enzyme and sequencing analysis. The most effective vectors were selected by RT-PCR and Western blotting after transfecting 293T cells. The best one was used to transform MHCC-97H cells which were selected with G418 to obtain positive colons. The changes of adhesion, invasion and FAK protein expression in MHCC-97H cells were studied. Results The restriction enzyme analysis and sequencing analysis demonstrated that shRNA had been inserted into vectors, and their sequences were the same as the design. It was indicated by RT-PCR and Western blotting that the silencing efficacy of IGF1R was 88%. The ability of adhesion and invasion significantly decreased after IGF1R silencing in MHCC-97H cells, and so was the expression of FAK protein. Conclusion IGF1R pGCsi-U6-Neo-GFP shRNA can significantly decrease the abilities of adhesion and invasion in MHCC-97H cells, and inhibit the expression of FAK protein.

[Objective] To investigate the protective effects of different extraction of Agaricus blazei Murrill on HaCaT cells damaged by UVB. [Methods] The cell viability which was detected by MTT method was used to determine the impact of different dose of UVB and different extraction of Agaricus blazei Murrill on normal HaCaT cells and HaCaT cells damaged by UVB. [Results] ①The HaCaT cells viability declined after irradiation with UVB, and it was a dose-dependent manner. The cell viability was 53.23%when the dose of UVB was 20mj/cm2. ②The water extraction at the concentration of 0.2~4mg·mL-1, aqueous extraction at the concentration of 0.1~2mg·mL-1 and crude polysaccharide at the concentration of 0.2~1mg·mL-1 could promote the proliferation of HaCaT cells, while aqueous extraction and crude polysaccharide inhibited the proliferation of HaCaT cells at higher concentrations.③The water extraction at the concentration of 1~4mg·mL-1, aqueous extraction at the concentration of 1~2mg·mL-1 and crude polysaccharide at the concentration of 0.5~1mg·mL-1 could enhance the cells viability which was irradiated by 20mj/cm2 UVB (P<0.01). [Conclusion] The different extraction of Agaricus blazei Murrill all could relieve the photo-damage caused by UVB, and it has the potential of anti-photoaging.

Arrhythmias, Cardiac ; complications ; physiopathology ; Atrioventricular Block ; complications ; physiopathology ; Coronary Circulation ; Humans ; Male ; Middle Aged ; Syncope ; complications ; physiopathology

Objective To study the effect of down-rdgulation of EF-1 alpha gene in prostate cancer cell line DU-145 on cancer cell migration and invasion by using RNA interference technique. Methods The prostate cancer cell line DU-145 was divided into three groups: the control group (untransfected with siRNA), randomly control group (randomly transfected with siRNA) and experimental group (transfected with EF-1 alpha siRNA). Localization of EF-1 alpha and its relationship with F-actin in cytoplasm were analyzed by immunofluorescence technique. Cancer cell migration and invasion capability of DU145 cells were studied by transwell technique in these three groups. Results EF-1 alpha expression in DU145 cell line was down-regulated by using RNA interference technique. EF-1 alpha was localized in cytoplasm and co-located with F-actin. The down-regualtion of EF-1 alpha did not change the F-actin distribution in cytoplasm. The cell migration and invasion study showed that after seeding 20×104 DU145 cells into the upper chamber of transwall for 12 hours, the cells collected in the lower chambers were (10.6±1.0)×104 in control group, (11.2±0.8)×104 in randomly control group and (3.9±0.6)×104 in experimental group. Compared with controls, the cancer cell migration and invasion capability was significantly inhibited to only 37.1% (t= 13.9, P<0.05) after the specific down-regulation of EF-1 alpha expression in DU145 cells. Conclusions The down-regulation of EF-1 alpha expression has negative impacts on prostate cancer cell migration and invasion. EF-1 alpha plays important roles in prostate cancer local invasion.

Objective To investigate the type distribution of urogenital Chlamydia trachomatis(Ct)among STD clinic outpatients in Guangxi Zhuang Autonomous Region, and to estimate the prevalence of Ct infection among the patients during posttreatment follow?up. Methods Urethral and cervical swabs were collected from male and female outpatients with confirmed urogenital Ct infection, respectively, in Institute of Dermatology of Guangxi Zhuang Autonomous Region. The patients with positive results in preliminary screening tests were followed up after treatment, and specimens were collected at follow?up visits. General and clinical information was also obtained from these patients. DNA was extracted from these samples by using the QIAxtractor instrument. Nested PCR was performed to amplify the major outer membrane protein A(ompA)gene for ompA typing, and to amplify CT046(hctB), CT058, CT144, CT172 and CT682 (pbpB) genes for high?resolution multilocus sequence typing (hr?MLST). Then, PCR products were sequenced, and ompA and MLST types of Ct were determined by sequence alignment and MLST analysis, respectively. The obtained MLST sequence types (STs) were compared with those from an Italian population by using the BioNumerics7 software, and a minimum spanning tree(MST)was generated. Results Totally, 44 and 6 Ct?positive specimens were collected at first visits and follow?up visits respectively. Among the 50 specimens, 42 underwent successful ompA typing and hr?MLST, and 7 ompA genotypes and 15 hr?MLST STs were identified, including 3 first reported STs. The distribution of STs of Ct isolates from Guangxi Zhuang Autonomous Region was significantly different from that from the Italian population. Among the 6 followed patients with posttreatment Ct infection, 3 were confirmed to be reinfected with Ct, and the other 3 failed to be diagnosed because of unsuccessful genotyping. Conclusion The genotypes of Ct strains isolated from STD clinic outpatients in Guangxi Autonomous Region were characteristic, and Ct reinfection occurred in some patients during follow?up.

Objective:To observe the stability of thymosin ?_1 microspheres under preparation conditions.Methods: Using HPLC method,we investigated the influence of different temperatures(-20 ℃, 6-8℃,60℃),sonification time(10-60 s) and pH values (pH 4.0,7.0 and 10.0) on thymosin ?_1 stability.Results: Under the conditions of frozen environment(-20℃) and refrigeration(6-8℃), thymosin ?_1 remained stable for 30 d.Thymosin ?_1 did not degrade in 60℃ water bath for 12 h or in phosphate buffer solution(PBS,pH 4.0) under 37℃ for 14 h.The sonification time within 60 s was found to be safe for the preparation.Conclusion: Thymosin ?_1 is stable under these preparation conditions.

Objective To study the elongation factor 1α(EF-1α) gene functions in prostate cancer cell line DU145 in the aspects of cell proliferation and clone formation by using the RNA interference technique. Methods DU145 cell lines were divided into control group, transfection control group transfected with scramble siRNA and experimental group transfected with EF-1α siRNA. After transfecting EF-1α siRNA into DU145 cell line, the down-regulation of EF-la expression in DU145 cell line was confirmed by Western blotting and immunofluorescence staining. Then, the cell proliferation and clone formation assays were carried on in these 3 groups of DU145 cells. Results Compared with controls, the specific down-regulation of EF-1α expression was achieved in experimental group only. Compared with control group, after the down-regualtion of EF-1α in DU145 cell line, the cell proliferation rate decreased from day 4 to day 7 after transfection by 45. 9%, 53. 5% , 35. 3% and 38. 1% , respectively(P<0. 05). The clone formation number in experimental group decreased by 67.0% (P<0. 01). Conclusions The down-regulation of EF-1α has a negative impact on prostate cancer cell proliferation and clone formation. EF-1α might be an appropiate targeting gene in prostate cancer targeting therapy.

A new method has been established for simultaneous determination of anions and cations in fertilizer sample by multimodal liquid chromatography with direct conductivity detection. An Acclaim Trinity P1 column based on nanopolymer silica hybrid technology with multimodal separation functional groups reversed-phase/anion-exchange/cation-exchange was used for the analysis. The chromatographic conditions were optimized and the effect ion of eluent on retention was discussed. Eight ions ( Li+, NH+4 , K+, HCOO-, NO-2 , Cl-, NO-3 and Br-) were separated and determined simultaneously by using 25 mmol/L CH3 COONa solution containing 50% acetonitrile at pH=5. 0 as mobile phase. The flow rate was 0. 50 mL/min and the temperature was 30 ℃. Under the optimum conditions, the linear ranges of the method were in the range of 0 . 5-200 mg/L for all the ions with correlation coefficient of 0 . 9997-0 . 9999 . Whereas the detection limits (S/N=3) were in the range of 0. 16-1. 72 mg/L and the relative standard deviations (RSD, n=9) were in the range of 1 . 3-2 . 5%. The method was applied to the determination of anions and cations in the fertilizer samples with satisfied results and the recoveries were in the range of 95 . 8%-103 . 8%.

To prepare thymosin alpha-1 (Talpha) loaded injectable sustained release microspheres and to evaluate its release behavior, bioactivities in vitro as well as its pharmacodynamics in vivo, Talpha1 loaded microspheres was prepared with poly ( lactic-co-glycolic acid) (PLGA) as carrier material by double emulsion (W/O/W) method. Physical and chemical properties of microspheres, such as mean diameter, The release behavior and its influencing factors were morphology and drug loading were evaluated. evaluated by HPLC determination. The bioactivity of Talpha1 in the course of encapsulation process and in vitro release ware evaluated by CCK-8 method. The ratio of CD4+/CD8+ in blood was determined with flow cytometry and the pharmacodynamics of Ta, loaded microspheres was evaluated by the change of CD4+/ CD8+. Microspheres with good shape and dispersive quality were prepared. The drug entrapment efficiency of two optimizing prescriptions containing 5% NaCl and 10% glucose as outer water phase were 87. 8% and 90. 2% , respectively. The cumulated release in one month is up to 90%. The bioactivity of Talpha was conserved with glucose as outer water phase, but in the course of in vitro release, the specific activity of Talpha in the microspheres decreased a little. Talpha microspheres can increase significantly the immunity of immuno-suppressed rats. Talpha can be encapsulated in injectable microspheres to yield one-month continuous release when using biodegradable polymers PLGA as carrier material, and this technique will have a favorable perspective in the near future.
Animals ; Biological Availability ; CD4-CD8 Ratio ; Delayed-Action Preparations ; Drug Carriers ; Drug Compounding ; methods ; Injections ; Lactic Acid ; chemistry ; Male ; Mice ; Mice, Inbred BALB C ; Microspheres ; Particle Size ; Polyglycolic Acid ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thymosin ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacokinetics