Objective To improve the knowledge about controlling plague in cadres, masses, and the medical staff in plague affected area in Dinghian county of Shaanxi province and to assess the efforts of health education activities. Methods In 2008, the education activities carded out by the government-related departments were investigated. The awareness of plague control and assessment was obtained through a written survey, questionnaire and interviews on clinic. Results Some education activities were carried out in plague area of Dingbian county, such as training, issuing educational materials, broadcasting plague scientific educational films and arranging knowledge grand prix. The rates of knowing plague clinic, epidemiology, prevention and the "three prohibitions and three alerts to report" were as follows: the cadres were 50.50%(101/200),63.69%(414/650),78.67%(118/150), the masses were 64.71% (66/102),87.91% (269/306),76.47% (78/102) and the medical staff were 64.18% (543/846) ,68.51%(322/470),67.02%(63/94). The passing rates of the cadres, the masses and the medical staff were 70.00% (35/50),92.16% (47/51),74.47% (35/47). Conclusions Health education can strengthen health consciousness of cadres and masses and improve the ability of the medical stalf on controlling sudden epidemic situation. Reinforcing plague control knowledge and training of medical staff are still important for health education in the future.

Objective To investigate the early clinical outcome of peripheral cutting balloon(PCB) in the management of peripheral vessel stenosis.Methods Thirteen patients with peripheral limb vessel stenosis, in which 4 stenoses in hemodialysis access and 9 at lower limb arteries,underwent angioplasty by PCB.For multiple stenosis in the same vessel,the distal one should be expanded firstly.The balloon pressure was controlled in the range of 8 atm to 10 atm(1 atm=10.108 kPa).All the patient were given continuous anticoagulant therapy after the procedure.Results All the procedure were carried out successfully on the 13 patients,and no serious complications occured.The symptoms did not recur in all patients after the procedure.The 5 months' follow-up angiography proved that no restenosis occurred in one patient with previous stenosis at the hemodialysis access.Conclusion The angioplasty with PCB was a safe and reliable procedure in management of the peripheral limb vessel stenosis.The early outcome is satisfying.

Objective To investigate the role of cathepsin G in photoaged fibroblasts. Methods Human fibroblasts were cultured and induced to premature senescence using UVA + MOP methods. Senescence-associated-β-galactosidase (SA-β-gal) stain was used to evaluate the positive rate of aged cells. The mRNA and protein expression of cathepsin G in photoaged fibroblasts were detected by real-time RT-PCR and Western blot techniques. Results Over 98 % induced cells presented a positive SA-β-gal straining. The expression of cathepsin G, detected by Western blot, was increased to (1. 70±0. 028) times of the control. And RT-PCR revealed that the synthesis of cathepsin G mRNA was also up-regulated to 1. 42±0. 09. Conclusion The results of our study demonstrates a significant correlation between photoaged fibroblasts and cathepsin G. The up-regulation of cathepsin G may play an important role in the damages of extracellular matrix and activation of MMPS in photoaged human skin.

OBJECTIVETo give some theory support of Cistanche tubulosa cultivation by searching dry matter accumulation and echinacoside content of C. tubulosa.

METHODDry matter accumulation content of C. tubulosa culturing in Huabei plain was analysed in different growth season of C. tubulosa. Echinacoside content was determined by HPLC.

RESULTDry matter accumulation of C. tubulosa showed "S" variation. Dry matter accumulation increased fastest in September among growing seasons. Dry matter amount was 138.58 g after C. tubulosa grew a year. Dry matter amount decreased significantly along with inoculation time retarded. Echinacoside content was 30.59% when C. tubulosa grew in 5 months, decreased guadully after that, and 9.76% in annual.

CONCLUSIONVariation rule of dry matter accumulation and echinacoside content was found in C. tubulosa that grew one year in Huabei plain.

Biomass ; China ; Cistanche ; chemistry ; growth & development ; Glycosides ; metabolism ; Plants, Medicinal ; chemistry ; growth & development ; Seasons

BACKGROUNDTo construct and express a phage display library of anti human lung cancer monoclonal antibody 5F-11.

METHODSImmunoglobulin variable regions (VH,VL) were amplified from 5F-11 hybridrom by RT-PCR. ScFv genes consisting of VH DNA and VL DNA joined together by a linker DNA were cloned into a phage vector pCANTAB5E. After 4 rounds of screening with lung adenocarcinoma cell line A2 as antigen, an enriched secondary phage display library was obtained.

RESULTSA recombinant phage display library with total of 8×10⁷ pfu/ml was established. Randomized clones from unselected library digested with BstNⅠ showed different patterns, however, those from selected library showed that phages with special pattern were enriched. Twenty-three out of 30 clones were found to respond strongly to A2 cell lines.

CONCLUSIONSThe ScFv of anti-lung adenocarcinoma monoclonal antibody 5F-11 can be successfully produced, which may be useful to widen the application of the antibody.

BACKGROUNDTo study the effects of extraneous p53 antisense RNA on malignant growth and sensitivity to cisplatin of human lung cancer cell line.

METHODS801D cell line with p53 deletion and mutation at 248 codon was selected as a parent cell line. An 1.8 kb human p53 full length cDNA was inserted into a mammalian expression vector PEGFP to construct a p53 antisense RNA recombined plasmid PEGFP-p53(AS) and GFP gene at plasmid was a report gene to monitor extraneous gene expression. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohitochemical stain of p53 monoclonal antibody. The inhibition growth efficacy of extraneous p53 in vitro was determined by clonogenic survival assay. Sensitivity of cells to cisplatin was examined with MTT assay. FCM analysis was performed to measure the effect of p53 antisense RNA on cell cycle.

RESULTSTwo cell lines, PEGFP-p53(AS)-801D and PEGFP-801D, were established after transfection of 801-D cells by lipofection and selection. Presence of extraneous p53 gene in PEGFP-p53(AS)-801D was proved by PCR and expression of extraneous p53 was estimated when green fluorescence in those cells was found out under the fluorescent microscopy. Mutated p53 protein in parent cell line 801D was positive and in PEGFP-p53(AS)-801D was negative with immunochemical stain. The inhibition rate of colony formation was 61% for PEGFP-p53(AS)-801D (P < 0.001). The sensitivity of PEGFP-p53(AS)-801D cells to cisplatin was increased. FCM analysis showed that the cell line was arrested at G1 phase.

CONCLUSIONSp53 mutation at 248 code plays an important role on malignant growth and resistance to cisplatin of human lung cancer cell line 801D. Malignant growth of cells with p53 deletion and mutation at 248 codon can be inhibited by extraneous p53 antisense RNA, and simultaneously the sensitivity to cisplatin is also increased.

OBJECTIVETo investigate the influence of fluorescent protein expression on the proliferation of murine NIH3T3 cells, so as to provide a theoretical basis for cell tracing technology.

METHODSNIH3T3 cells were cultured in vitro, and were randomly divided into control, pLEGFP-N1 (with transfection of pLEGFP-N1 retroviral vector), pEGFP-N1 (with transfection of pEGFP-N1 vector) and pDsRed2-C1 (with transfection of pDsRed2-C1 vector) groups. Then the cells were screened by G418 for 3 weeks. The changes in cell adhesive rate were observed and the population doubling times was determined by growth curve.

RESULTSThere was obvious fluorescent protein expression in the transfected NIH3T3 cells after G418 selection, and the highest percentage of labeled NIH3T3 cells was found in pLEGFP-N1 group. The population doubling time in pDsRed2-C1 (40.3+/-0.7 h) , PEGFP-N1 (39.6 +/- 0.6 h) and pLEGFP-N1 (36.5 +/- 0.7 h) groups was evidently longer than that in control (27.9 +/- 0.6 h, P < 0.01), with high adhesive rate in each group.

CONCLUSIONThe expression of fluorescent protein exhibited some inhibitory effect on the proliferation of NIH3T3 cells in vitro. Since the inhibitory effect by retroviral vector was weaker compared with eukaryotic vector, it should be the first choice for fluorescent protein labeling during cell transplantation.

Animals ; Cell Culture Techniques ; Cell Proliferation ; Green Fluorescent Proteins ; biosynthesis ; Mice ; NIH 3T3 Cells ; Transfection

BACKGROUNDTobacco-specific 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is the most important carcinogen in cigarette. Models induced by NNK are widely used in investigations about the mechanisms of pulmonary neoplasia and chemoprevention studies. The aim of this study is to explore the pulmonary precancerous lesions induced by NNK and its possible mechanisms.

METHODSFifteen Wistar rats were divided into two trial groups, in which the high-dose group was instilled with iodized oil including 10 mg (50 mg/kg) NNK into the left lower lobar bronchus, and the low-dose group received 5mg ( 25mg/kg) NNK. Another 15 Wistar rats were instilled only with iodized oil as control group. All rats were examined immediately after instillation and followed up periodically by pulmogram. The pulmonary tissues of rats were pathologically examined, and the expression of AE1/AE3, PCNA and p53 was detected by immunohistochemical method.

RESULTSThe pulmograms showed that the iodized oil localized at the bottom of left lobe and disappeared 107 days later. In trial group, 10 of 15 rats (67%) had nodus at the bottom of left lobe. All of rats in trial group (15/15) displayed atypical hyperplasia in alveolar region, showing single or multiple layers of proliferative epithelial cells along intact alveolar septa with irregular and non-discrete margins of lesion, but continuous alveolar spaces were not obliterated by proliferative epithelial cells. Ten of 15 rats in trial group showed severe atypical hyperplasia of glandular epithelium with occasional infiltrating to muscular layer. All of those atypical hyperplasia cells showed positive AE1/AE3 expression. The positive rate of PCNA was 90% (9/10) and 100% (5/5) in low-dose group and high-dose group respectively, which was significantly higher than that in control group (13%, 2/15) (P=0.000, P=0.001). The positive rate of p53 expression was 50% (5/10) and 60% (3/5) in low-dose group and high-dose group respectively, which was significantly higher than that in control group (0) (P=0.005, P=0.009). However, there was no remarkable difference in PCNA and p53 expression between low-dose group and high-dose group (P > 0.05).

CONCLUSIONSTransbronchial instillation of iodized oil including tobacco-specific NNK can induce pulmonary lesions as atypical hyperplasia of alveolar cell and glandular epithelium in Wistar rats. This model can be used in experimental studies about tobacco-related lung cancer.

OBJECTIVETo prepare and observe the physicochemical properties of scaffold materials of heterogeneous deproteinized tissue-engineered bone.

METHODSDeproteinized bone was made through a series of physicochemical treatments in pig ribs and analyzed with histological observation, scanning electron microscopy, infrared spectrum, X-ray diffraction and energy dispersive analysis, Kjeldahl determination and mechanics analysis.

RESULTSInterstitial collagen fiber was positive and mucin was negative in deproteinized bone, but, both were positive in fresh bone. Deproteinized bone maintained natural pore network. Its pore size was 472.51 micromolar+/-7.02 micromolar and the porosity was 78.15%+/-6.45%. The results of infrared spectrum showed that collagen was present in deproteinized bone. Both fresh and deproteinized bone had curve of hydroxyapatite. The Ca/P ratios were 1.71+/-0.95 and 1.68+/-0.76 (P larger than 0.05), and the protein contents were 26.6%+/-2.23% and 19.1%+/-2.14% (P less than 0.05) in fresh and deproteinized bone, respectively. There was no significant difference of destruction load under compression and maximal destruction load between fresh and deproteinized bone (P larger than 0.05). The elastic modulus was higher in deproteinized bone than that in fresh bone (P less than 0.05).

CONCLUSIONSPhysicochemical properties and mechanic strength of deproteinized tissue-engineered bone meet the demands of ideal scaffold materials. But, its immunogenicity should be observed through further experiments for its clinical applications.

Animals ; Biomechanical Phenomena ; Bone and Bones ; chemistry ; physiology ; Hydroxyapatites ; Materials Testing ; Swine ; Tissue Engineering

OBJECTIVETo evaluate the biological safety of manufactured heterologous deproteinized bone and to provide an experimental basis for clinical applications.

METHODSDeproteinized bone (10 mm) and leaching liquor were made from pig ribs with a series of physical and chemical methods, then were evaluated through acute and subacute toxicity test, hemolysis test, pyrogen test, intracutaneous test, intramuscular implantation test and cytotoxity test.

RESULTSNo obvious toxicity, hemolysis, pyrogenic characteristics, skin irritation, inflammatory reaction after intramuscular implantation and cytotoxity were observed.

CONCLUSIONSThe heterologous deproteinized bone has good biological safety and meets all the demands of scaffold material for tissue engineering.

Animals ; Bone Transplantation ; Cytotoxicity Tests, Immunologic ; Hemolysis ; Mice ; Rabbits ; Tissue Engineering ; Transplantation, Heterologous