4-Hydroxycoumarins ; poisoning ; Child ; Female ; Humans ; Lower Extremity ; blood supply ; Thrombosis ; etiology

The aim of this paper is to report the study on gene silencing efficiency of siRNA targeted against mouse VEGFR2 (siVEGFR2) in vitro mediated by polyethyleneimine (PEI) and its anti-tumor effect in vivo. CY3-labeled siRNA was compounded into PEI and transfected into MS1 cells. Confocal microscopy was used to image the subcellular distribution of siRNA in MS1 cells. Semi-quantitative RT-PCR and Western blotting were used to evaluate VEGFR2 gene silencing induced by siVEGFR2/PEI complexes. A tumor-bearing nude mice model was established to compare the anti-tumor effect after delivered by local and systemic routes. siVEGFR2/PEI complex-transfected cells exhibited much fluorescence in cytoplasm with no evidence of nuclear accumulation. The expression levels of VEGFR2 mRNA and protein in PEI-transfected cells were significantly down-regulated compared with that in blank group, the silencing efficiency were 28.2% and 23.6% respectively. The tumor sizes in mice intratumorally injected with siVEGFR2/PEI complexes (189.429 +/- 17.562 mm3) were reduced definitely compared to that in mice injected with siVEGFR2/PEI complexes via vein route (315.507 +/- 20.491 mm3), or to saline groups (365.844 +/- 20.713 mm3). The study demonstrated that PEI could effectively transfect siRNA into cells and silence the VEGFR2 gene expression. Intratumoral delivery is more suitable for non-targeted modified PEI/siRNA complexes to inhibit the tumor growth in vivo. The present data lay a solid foundation to further study on the gene silencing mechanism for PEI-medicated RNAi and its anti-tumor efficiency in vivo.

BACKGROUND: The death of aging cells is virtually apoptosis. To a certain extent, it can be interpreted as a series of results of gere activities.Therefore, the inhibition of oncogene's expression can lengthen the life span of cells and delay aging of brain tissues.OBJECTIVE: To study the effect of abstragulus mongholicus bung (AMB) on apoptosis of nerve cells and the expression of relevant gene in aging mice brain.DESIGN: Completely randomized design and controlled experiment.SETTING: Department of Biochemistry, School of Basic Medical Sciences of Jiamusi University.MATERIALS: The experiment was conducted at the Experimental Animal Center and Biochemical Laboratory of Jiamusi University from December 2003 to May 2004. Totally 24 healthy Kunming mice were recruited in this study. There were 8 two-month-old mice (young group) and 16 twelve-month-old mice. All the 16 mice were randomized into abstragulus mongholicus bung group and old control group with 8 in each group.METHODS:① AMB group:Mice in AMB group received gastric gavage was provided by the Department of Traditional Chinese Medicine of the First Hospital Affiliated to Jiamusi University, and evaluated by Jiamusi Drug Inspection Bureau. Water decoction was prepared with 2 kg/L raw materials. Mice in old control group and young group were filled with lukewarm boiled water.② All the animals were treated as above for 30 consecutive days before put to death. Their brains were taken out immediately and the middle parts of the brains were removed to fix with neutral formaldehyde. The remaining brain tissues were made into mitochondria suspension. Content of manganese superoxide dismutase (Mn-SOD) and malondialdehyde (MDA) was determined with xanthosine oxidase method and TBA chemical colorimetry. Apoptotic cells (cells with yellow nuclei were positive ones) were assayed with in situ end-labeling (ISEL) and expression of bcl-2 gene was assayed with immunohistochemical method. The cells stained brown were positive ones. A total of 400 cells were counted under the 400× microscope. We graded the samples according to the percentage of the positive cells: the number of positive cells ＜ 5% -; 5%-10％ +; 11％-50％ ++; ＞ 51％ ().③ Grade and quantitative data were compared with rank sum test and t-test.MAIN OUTCOME MEASURES: Effects of AMB on the rate of neu-ronal apoptosis, the activity of Mn-SOD, the concentration of MDA in mitochondria, and the intensity of the expression of bcl-2 gene.RESULTS: Totally 24 mice entered the final analysis.① Content of MnSOD was higher in young group and AMB group than in old control group (P＜0.05).② Concentration of MDA and apoptotic rate in young group and AMB group were lower than those in old control group (P ＜ 0.01).③Expression of bcl-2 gene was significantly different in young group and AMB group from that in old control group (P ＜ 0.01).CONCLUSION: AMB is found to be able to obviously inhibit neuronal apoptosis in aging mice brain by affecting the activity of Mn-SOD, the concentration of MDA and the expression of bcl-2.

Objective To dynamically observe the changes of the plasma superoxide dismutase(SOD) activity and malondialde-hyde(MDA) content of rabbits with hemorrhagic shock in high latitude .Methods 28 rabbits were randomly divided into 4 groups :0 .9% NaCl group ,7 .5% NaCl group ,Voluven(6% HES 130/0 .4) group and control group .The plasma SOD activity and MDA content were measured before shock .At 30 minutes of shock and 30 minutes later of volumetric resuscitation .Then the changes of MAP were observed .Results There were no static difference among 4 groups in the levels of SOD and MDA before shock ;at 30 minutes of shock ,the SOD activity was reduced signicantly and the MDA content was increased in each group (P<0 .01) .Afer fluid infusion there were no obvious change in the plasma levels of SOD and MDA compared with those of shock in 0 .9% NaCl group and in 7 .5% NaCl group(P>0 .05) ,whereas in Voluven group ,compared with those in 0 .9% NaCl and 7 .5% NaCl group the SOD activity was elevated signicantly and the MDA content was decreased (P<0 .01) .Conclusion Voluven can be used in scan-venging oxygen free redicals by recovering the plasma SOD activity in rabbits with hemorrhagic shock in high latitude .

Objective To study the severity of systemic inflammatory response to type B epidemic encephalitis and blood tumor necrosis factor complicated with respiratory failure.Methods Fourteen children with type B epidemic encephalitis were divided into two groups;6 cases in group one with respiratory failure were given mechanical ventilation and the other 8 cases had normal respiration.Breat-hing pattern,respiratory rate,GCS scores,daily dosage of sedatives were observed before endotracheal intubation;while heart rate,temperature,WBC,CRP,TNF were measured.Results Patients in respiratory failure group were given a small dosage of Luminal;there was no significant difference in GCS between two groups treated with compound Wintermin.The heart rate,temperature,WBC,CRP,TNF levels in respiratory failure group were significantly higher than those in normal respiratory group.Conclusions The severity of systemic inflammatory response is more severe in epidemic type B encephalitis with respiratory failure than the controls.Significant increase in blood TNF may be an important factors to cause peripheral respiratory failure.

Objective To analyze the false-positive results of Treponema pallidum antibody caused by 3 different assay in comparison with Treponema pallidum hemagglutination assay (TPHA).Methods Research group included 3957 clinically asymptomatic syphilis patients,and control group was 344 outpatients with sex-transmitted diseases (STD).The serum samples from the patients who were TPHA-positive were tested in parallel by enzymeimmunoassay (EIA) and syphilis toluidine red untreated serum test (TRUST).Western blot (WB) was performed as confirmatory test.Results In the clinically asymptomatic patients,60 were TPHA-positive.Among them 57 were confirmed by western blot assay,and 1 was false-positive and 2 were borderline in WB.Of the 60 TPHA-positive patients,53 were positive in EIA and 23 were positive in TRUST.In STD patients 40 were TPHA,WB and EIA-positive but 32 were TRUST-positive.Conclusions The results of TPHA and EIA were consistent for diagnosis of syphilis patients who may suffer from previous or latent infection.

Objective To observe and analyze the occurrence and related factors of pulmonary fibrosis caused by dust exposure in Shenzhen city in 2014 ,and to explore the preventive measures .Methods 64 cases of patients with pneumoconiosis from January 1st ,2014 to December 31st were selected as the study objects ,the factors affecting the development of pneumoconiosis and the re‐lated factors of complications were compared and analyzed ,the preventing measures were discussed .Results the proportion of pneumoconiosis stage Ⅱ and stage Ⅲ and the incidence of complications were increased with the working age of dust‐exposed work ,the differences were statistically significant(P<0 .05) .Compared with patients who used anti‐dust respirator and were trea‐ted in time ,the patients used common respirator and not treated in time had higher proportion of pneumoconiosis stage Ⅱ and stageⅢ and the incidence of complications ,the differences were statistically significant(P<0 .05) .The incidence of complications associ‐ated with pneumoconiosis such as tuberculosis ,pulmonary emphysema ,pulmonary heart disease were higher in patients with more than 20 working years than those with 10- <16 and 16-20 years ,the differences were statistically significant(P<0 .05) .Conclu‐sion Working years of dust‐exposed work ,personal protective measures ,timely treatment are major influence factors for the occur‐rence and development of pneumoconiosis and related complications .Strengthening the engineering and technical measures for re‐ducing dust in dust‐exposed work ,personal protection and health education ,regular physical examination are important means to prevent the occurrence and development of pulmonary fibrosis caused by dust‐exposed work .

OBJECTIVE: To investigate the functional mechanism of metadherin (MTDH), hypoxia-inducible factor-1 alpha (HIF-1α) and transketolase-like gene 1 (TKTL1) and their interaction with each other in laryngeal carcinoma development. METHOD: Thirty laryngeal carcinoma samples and 20 samples of para-carcinoma tissue were taken from the patients received operation treatment. The expression levels of MTDH, HIF-1α and TKTL1 were detected in all the samples by SP immunohistochemical methods. The data were analyzed by the SPSS13.0 statistical software. RESULT: The positive expression rate of MTDH, HIF-1α and TKTL1 in the 30 cases of laryngeal carcinoma was 56.67%, 60.00% and 63.33%, respectively, which was 20.00%,10.00% and 15.00% respectively in the para-carcinoma tissue, the difference of the positive expression rate of the tested objects between these two groups was statistically significant (P < 0.05 or P < 0.01). Of the 30 cases of laryngeal carcinoma, the positive expression rate of MTDH, HIF-1α and TKTL1 in the cases with lymphnode metastasis was 84.62%, 84.62% and 79.62%, respectively, compared with the rate in those without lymph nodes metastasis, which was 35.29%, 41.18% and 35.29%. The difference of the positive expression rate of the tested objects between these two groups was statistically significant (P < 0.05). In the tissue of poorly differentiated squamons cell carcinoma verified by pathology, the positive expression rates of MTDH and HIF-1α was 73.68% and 84.21%, respectively, while that in the tissue of well differentiated squamous carcinoma was 27.27% and 18.18%. The difference of the positive expression rate of the tested objects between these two groups was statistically significant (P < 0.05 or (P < 0.01). The positive expression rate of TKTL1 in laryngeal carcinoma at stage I-II was 25.00% and that at stage III-IV was 72.22%, the difference between which was statistically significant (P < 0.05). The positive expression rate of HIF-1α in laryngeal carcinoma at stage I-II was 33.33% and that at stage III-IV was 77. 78%, the difference between which was statistically significant (P < 0.05). The expression of MTDH, HIF-1α and TKTL1 showed no relationship with age,smoking amount or the tumor location (P > 0.05). The Spearman's rank correlation analysis showed that the positive expression rates of MTDH and HIF-1α in laryngeal carcinoma had positive correlation (r = 0.384, P < 0.05); the positive expression rates of TKTL1 and HIF-1α in laryngeal carcinoma had positive correlation (r = 0.508, P < 0.01); But there was no significant correlation between the positive expression rates of MTDH and TKTL1 in laryngeal carcinoma (r = -0.107, P > 0.05). CONCLUSION It suggested that these three proteins may have close relationship with the occurrence, invasion and metastasis of the laryngeal carcinoma. MTDH and TKTL1 may be expected to be new clinical targets for laryngeal carcinoma treatment and it could offer theoretical basis for the prognosis of the laryngeal carcinoma.
Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Cell Adhesion Molecules ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Laryngeal Neoplasms ; diagnosis ; metabolism ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; Transketolase ; metabolism

OBJECTIVE: To explore the expression of phospholipase Cgamma-2 (PLCgamma-2), lipoxygenase-12 (12-LOX) and arachidonic acid (AA) in laryngeal squamous cell carcinoma and to study the the relationship between lipid metabolism and laryngeal squamous cell carcinoma. METHOD: In 30 cases of carcinoma tissue and peritumoral laryngeal mucosa tissues (confirmed to be normal laryngeal tissues by pathology), immunohistochemical method (Streptavidin-peroxidase method, SP method) was used for the detection of expression of PLCgamma-2 and 12-LOX, and gas chromatography/mass spectrometry (GC/MS) for the content of the arachidonic acid in carcinoma tissue and peritumoral normal laryngeal mucosa tissues. RESULT: The positive rates of PLCgamma-2 and 12-LOX in carcinoma tissue were higher than in peritumoral normal laryngeal mucosa tissues with statistically significance differences (P < 0.05). The content of arachidonic acid was lower in carcinoma tissue than in peritumoral normal laryngeal mucosa tissues with statistically significance difference (P < 0.05). The positive expressions of PLCgamma-2 and 12-LOX were closely correlated to tnm stage, histological differentiation and lymph node metastasis (P < 0.05). The content of arachidonic acid had no significant correlations with tnm stage, histological differentiation and lymph node metastasis (P > 0.05). Both the expression of PLCgamma-2 and 12-LOX and the content of arachidonic acid had no statistically significant correlation with age (P > 0.05). CONCLUSION PLCgamma-2, AA and 12-LOX play important roles in laryngeal squamous cell carcinoma. It may be meaningful to the treatment of laryngeal carcinoma by suppressing this passway.
Aged ; Arachidonate 12-Lipoxygenase ; metabolism ; Arachidonic Acid ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Head and Neck Neoplasms ; metabolism ; pathology ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Phospholipase C gamma ; metabolism ; Squamous Cell Carcinoma of Head and Neck

OBJECTIVE: The purpose of this study was to investigate the expression of PKM2 and HIF-1α in laryngeal squamous cell carcinoma and to analyze their correlation in laryngeal squamous cell carcinoma. METHOD: Total 37 laryngeal carcinoma samples and para-carcinoma tissues were taken from the patients who accepted operation treatment in the Second Hospital of HeBei Medical University from 06/2013 to 06/2014. The protein expression levels of PKM2 and HIF-1α were detected with SP immunohistochemical methods. The data were analyzed by the SPSS 13.0 statistical software. RESULT: The positive expression of PKM2 in laryngeal carcinoma tissues and adjacent tissues were 62.16% and 13.15%. The difference was statistically significant (P < 0.01). The positive expression of HIF-1α in laryngeal carcinoma tissues and adjacent tissues were 64.86% and 21.62%. The difference was statistically significant (P < 0.01). The positive expression of PKM2 and HIF-1α in well differentiated laryngeal squamous cell carcinoma were both 47.83%, while in medium and poorly differentiated laryngeal squamous cell carcinoma were 85.71% and 92.86% respectively. The difference was statistically significant (P < 0.05). The positive expression of PKM2 and HIF-1α in patients with lymph metastasis were 90.00% and 100.00% respectively, 51.85% in those without lymph metastasis, the difference was statistically significant (P < 0.05). The rate of HIF-1α positive expression in I-II stage was 53.85%, 90.91% in III-IV stage. The difference between the two groups was statistically significant (P < 0.05). The expression of PKM2 and HIF-1α had no relationship with the age and smoking (P > 0.05). The expression of PKM2 was positively related with HIF-1α in laryngeal squamous cell carcinoma (P < 0.01). CONCLUSION The expression of PKM2 and HIF-1α are related with the proliferation, invasion and metastasis of laryngeal squamous cell carcinoma. It provides a certain theoretical basis for laryngeal cancer diagnosis and screening to measure the expression of PKM2 and HIF-1α as biological indicators.
Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Carrier Proteins ; metabolism ; Head and Neck Neoplasms ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Laryngeal Neoplasms ; metabolism ; Lymphatic Metastasis ; Membrane Proteins ; metabolism ; Squamous Cell Carcinoma of Head and Neck ; Thyroid Hormones ; metabolism