OBJECTIVETo investigate the effect of a phosphorothioate antisense oligodeoxynucleotide "ASOND" combined with cis-Diamminedichloroplatinum (DDP), 5-fluorouracil (5-FU) and adriamycin (ADM) respectively on inhibiting the proliferation of HepG2 cells.

METHODSA phosphorothioate antisense oligodeoxynucleotide (5'-ACTCACTCAGG CCTCAGACT-3') targeted to human telomerase reverse transcriptase (hTERT) mRNA, which named cantide, was synthesized. ASODN was transfected into HepG2 by lipofectin. And cell growth activity was evaluated by MTT assay. SAS software and Jin Zhengjun Method were used to evaluate the interaction of ASODN and these chemotherapeutic drugs.

RESULTSCombination treatments with 0.1micromol/L ASODN reduced the IC50 of DDP, 5-FU and ADM from 1.07, 4.15 and 0.29microg/ml to 0.25, 1.52 and 0.12microg/ml respectively. The inhibitory ability of combination treatments on HepG2 cells was higher than that of these drugs alone (F=66.92, 25.96, 8.56, P<0.001). And synergism (Q>or=1.15) was observed at the lower concentration of DDP (

CONCLUSIONASODN may enhance therapeutic effectiveness of chemotherapeutic drugs in human hepatocellular carcinoma cells.

Antineoplastic Agents ; administration & dosage ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; DNA-Binding Proteins ; Doxorubicin ; administration & dosage ; Drug Synergism ; Fluorouracil ; administration & dosage ; Humans ; Liver Neoplasms ; drug therapy ; Oligodeoxyribonucleotides, Antisense ; administration & dosage ; Telomerase ; antagonists & inhibitors ; genetics

Objective:To study the expression of coxsackievirus group B3 gene fragment encoding VP1 in procaryon and to explore its application.Methods:Stablie expression of VP1 gene of CVB3 in E.coli was obtained.The expressed protein was purified by NAT chromatography and its immunoactivity was identified by indirect ELISA.Results:The expressed product of VP1,similar to native protein antigen of CVB3,could strong bind with the mouse's antibody serum against CVB3(polyclonal antibody).Irrelevant monoclonal antibody as contrast presents negative activity. Using the expressed VP1 product,we have had a special IgM ELISA for the patient's serum of the clinical acute viral myocarditis.The result was same with the cellular protein antigens of the tissue-cultivated CVB3.Conclusion:The protein antigen CVB3-VP1 which is obtained by the method of gene engineering has character of high product, and its immunoactivity after being purified was basically unchanged. At present, this kind of antigen is difficult to be obtained from the viral cullture medium and a potent hazard for being infected by this virus may take place in such manipulateion. By the method of gene engineering we can obtain antigenic VP1 of CVB3 and use as immuneogen for the detection of serum antibody,by which to provide reliable test reference for the early-stage diagnosis and clinical therapy of the acute myocarditis.

OBJECTIVETo observe the therapeutic effect of Qingfei Huatan Quyu method (QHQ, a Chinese medicinal therapy for clearing Fei-heat and dissolving phlegm-stasis) combined with hormone-antibiotic therapy (HAT) on radiation pneumonia (RP).

METHODSEighty-one patients with RP were randomized into two groups, 41 patients in the control group and 40 in the treatment group were treated with HAT alone and HAT combined with QHQ respectively for 21 days. The severity of RP was evaluated before and after treatment according to the criteria of the radiation therapy oncology group. The effect on TCM symptoms and chest roentgenogram, as well as on plasma levels of interleukin-6 ( IL-6) and transform growth factor-beta (TGF-beta) were detected.

RESULTSAfter treatment, number of patients with RP graded as 0, 1, 2, 3, and 4 in the treatment group was 23, 10, 4, 2, and 1, respectively, while in the control group, 14, 9, 11, 4, and 3, respectively. The combined therapy showed effects in improving RP grading (P <0.01) and TCM syndromes were superior to those of HAT respectively (P < 0.05). Besides, levels of IL-6 and TGF-beta were lowered after treatment in the treatment group, showing a significant difference to those in the control group (P <0.05).

CONCLUSIONQHQ combined with HAT has a definite therapeutic effect on RP. It could efficiently decrease the plasma levels of IL-6 and TGF-beta in patients with RP.

Anti-Bacterial Agents ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Interleukin-6 ; blood ; Medicine, Chinese Traditional ; Radiation Pneumonitis ; drug therapy ; Transforming Growth Factor beta ; blood

0.05),but a significant difference at weeks 4,8,and 12 between two groups(P

BackgroundIron-containing foreign body trapped in the eyeball wall without affecting the opticus occurs occasionally in clinic. Operation always is performed in an attempt to avoid the deposition of rust in different tissues of the eye-balls. However,a few animal experimental studies showed that a small foreign body does not affect the retina and opticus in the period of three months. The question of whether surgery needs to be carried out is worth discussion. ObjectiveThe aim of this study was to evaluate the influence of posterior intrascleral iron foreign body on the rabbit retina and opticus. MethodsTwelve healthy adult Japan flap-eared white rabbits were randomly divided into two groups. Medium carbon iron with rust or without rust( size of 2. 0 mm × 1. 0 mm×0. 2 mm) were implanted into the posterior sclera of the left eye to create the animal model with iron foreign body in the eyeball wall. The cornea, anterior chamber, crystalline lens, vitreous and fundus of the rabbits were observed under a slit lamp microscope 1weekbeforeoperationand 1week, 2weeks, 1monthand 3months after operation.Flash electroretinogram(F-ERG) and flash visual evoked potential (F-VEP) were recorded at the time points mentioned above. All the rabbits were sacrificed and the eye balls were extracted at the end of the experiment, and the position of the iron foreign body was determined. The histopathological examinations of the retina and opticus were performed under the light microscope. This experiment complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThere were no statistical differences for the a-wave amplitude of F-ERG among different time points( F =1. 885,P =0. 129 ) and different treatment groups ( F =1. 188, P =0. 340 ), as with the ERG b-wave amplitude ( time: F =2. 73, P =0. 064 ; group : F =1. 114, P =0. 367). The differences in the latencies of F-VEP N1-wave were insignificant among the different time points( F =1. 605, P =0. 263 ) as well as various groups ( F=1. 556, P =0.314 ), and those of F-VEP P1 -wave were not evidently changed ( time: F =2. 329, P =0. 092 ; group : F =2. 186, P =0. 103 ). No correlations were seen between the time factor and grouping factor ( P ＞ 0. 05 ). There was no apparent siderosis bulbi change during the follow-up duration. No morphological abnormality in the retina and optical nerve was found under the light microscope. At the end of the experiment,intrascleral iron foreign body was wrapped by surrounding tissue in a stable condition. Conclusions The small posterior intrascleral iron foreign body, whether it is oxidized or not, does not produce distinctive functional or pathological damage on retina and opticus in the short term.

Objective To investigate the effect of mesenchymal stem cells (MSC) on the activity of nuclear factor (NF)-?B in rats with myocardial infarction.Methods MSC were isolated from SD rats (120—150 g in weight).SD rats (180—200 g in weight) were subjected to MI by left coronary artery occlusion,and were allo- cated into three groups randomly:1)sham group (without ligation of the artery,n=8);2)injection of PBS solu- tion (n=8);3)injection of MSC (n=8).MSC or PBS solution was injected into myocardium from epicardium instantly after MI models were established.Four weeks after transplantation,cardiac function was evaluated u- sing physiological recorder.Western blot were performed to investigate the nuclear factor-? activity.The ex- pressions of tumor necrosis factor (TNF)-? and interleukin (IL)-6 were evaluated by RT-PCR and Western blot. Results 1)Mortality was 20%(2/10) in sham group,33.3%(4/12) in PBS group and 20%(2/10) in MSC group with no statistic differences between them(P=0.646).2) Hemodynamic measurements showed that MSC trans- plantation caused significant improvement in cardiac function,comparing with MI+PBS group.3) MSC inhibi- ted the activities of NF-?B in myocardium and down-regulated the expression of TNF-? and IL-6 in mRNA and protein level.Conclusion Transplantation of MSC improved cardiac function in MI rats,which may partly at- tribute to their immuno-inflammatory regulation mechanism.

OBJECTIVEBy using Benchmark Dose (BMD) approach to explore the relations among drinking water fluoride, urine fluoride, serum fluoride and dental fluorosis; and to evaluate the significance of urine fluoride and serum fluoride in control and prevention of endemic fluorosis.

METHODS512 children (290 in Xinhuai Village, 222 in Wamiao Village) aged 8-13 years were recruited in the study. Epidemiological methods were used to investigate the prevalence of dental fluorosis, and the levels of urine fluoride, serum fluoride, and drinking water fluoride in superficial well. The children were divided into six subgroups by the concentration of fluoride in drinking water: < 0.5 mg/L, 0.5-mg/L, 1.0-mg/L, 2.0-mg/L, 3.0-mg/L and > or = 4.0 mg/L.

RESULTSThere was significant dose-response relationship between the drinking water fluoride and the prevalence of dental fluorosis or the prevalence of defect dental fluorosis. The BMDLs (Benchmark Dose Lower Bound) were 1.01 and 1.30 mg/L, respectively. Urine fluoride and serum fluoride also had significant dose-response relationship to the prevalence of dental fluorosis or defect dental fluorosis. The correlation coefficient between drinking water fluoride and urine fluoride was 0.717, and it was 0.855 between drinking water fluoride and serum fluoride, and 0.617 between urine fluoride and serum fluoride.

CONCLUSIONSThe currently national standard of fluoride in drinking water in China is safe and reasonable. As a biological monitoring index, the levels of fluoride in serum may be more useful than that in urine in the control and prevention of endemic fluorosis.

Adolescent ; Child ; China ; epidemiology ; Environmental Monitoring ; Epidemiological Monitoring ; Female ; Fluorides ; analysis ; blood ; urine ; Fluorosis, Dental ; epidemiology ; Humans ; Male ; Prevalence ; Water Supply ; analysis ; standards

[Objective] To observe the specific immune responses induced by the recombinant major outer membrane protein (rMOMP) vaccine against Chlamydia trachomatis E serotype in rhesus monkeys.[Methods] Six rhesus monkeys were equally divided into three groups:adjuvant and protein group vaccinated with purified rMOMP and Freund's adjuvants,adjuvant group immunized with Freund's adjuvants only,and control group immunized with phosphate buffer.All the rhesus monkeys were intramuscularly immunized in the triceps brachii for 3 times at a 2-week interval.Two weeks after the last vaccination,serum,vaginal wash and venous blood samples were collected from the rhesus monkeys,and lymphocytes were isolated from the blood samples.Enzyme linked immunosorbent assay (ELISA) was performed to determine the specific IgG antibody and interferon level in sera and secretory IgA (sIgA) level in wash samples,and methyl thiazolyl tetrazolium (MTT) assay to evaluate the proliferation of lymphocytes after stimulation with Chlaraydia trachomatis serotype E elementary bodies.Delayed hypersensitivity was observed in rhesus monkeys challenged by inactivated Chlamydia trachomatis serotype E elementary bodies.In vitro antibody neutralization assay was conducted with the serum from rhesus monkeys.Indirect immunofluorescenee was used to detect Chlamydia trachomatis in exfoliative vaginal cells from rhesus monkeys from week 1 to 10 after challenge with Chlamydia trachomatis.Data were statistically analyzed by using one-way analysis of variance and least significant difference (LSD) test with the SPSS 14.0 software.[Results] The adjuvant and protein group differed statistically from the adjuvant group and control group in the serum level of specific IgG antibody (1.718 ± 0.213 vs.0.841 ± 0.315 and 0.791 ±0.437,both P＜ 0.05),interferon ((1086 ± 121.730) ng/L vs.(409 + 53.440) ng/L and (162 ± 48.046) ng/L,both P＜ 0.05),lymphocyte proliferation index (7.012 ± 1.026 vs.4.473 ± 1.850 and 1A26 ± 1.104,both P＜0.01 ) and the diameter of nodus in delayed hypersensitivity assay ( ( 1 1 ± 2.134) mm vs.(3 ± 0.914) mm and 0,both P ＜ 0.01 ).After attack,the exfoliative cells kept positive for Chlamydia trachomatis in the adjuvant and protein group from week 1 to 5,and in the other 2 groups from week 1 to 10,but were negative in the adjuvant and protein group from week 6 to 10.[Conclusion] The rMOMP vaccine can induce a specific,protective,humoral and cellular immune response against Chlamydia tracbomatis in rhesus monkeys.

OBJECTIVESTo examine the activities of transcription factors (TFs) in human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to identify the TFs associated with HCC metastasis.

METHODSTranscription factor activity profile of Hep3B, MHCC97L and MHCC97H, three HCC cell lines with different metastatic potentials, were examined using protein/DNA array. Electrophoretic mobility shift assays (EMSA) and Western blot were used to confirm the results obtained by protein/DNA array.

RESULTSFrom a total of 345 screened TFs, 7 activity differential TFs were found, of which 5 showed increased activity, including p53, hypoxia inducible factor-1 alpha (HIF-1alpha), signal transducer and activator of transcription 3 (Stat3) and Sp1, and 2 showed decreased activity including Rb and Smad3.

CONCLUSIONThe abnormal functioning of transcription factors is closely associated with HCC metastasis. Our present findings could be of help in expanding our understanding of the mechanism of HCC metastasis and identify new predictive biomarkers and therapeutic targets.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; DNA Fingerprinting ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Metastasis ; Protein Array Analysis ; Transcription Factors ; classification ; genetics ; metabolism

OBJECTIVETo observe the effects of the fat flap tissues after delay operation on free fat-graft survival rate and duration.

METHODSThe delay operation of fat flaps was performed in the inguinal region of a rabbit. Expression of VEGF was assayed using Elisa method after 12 hours of flap delay. The fat flaps were harvested and cut into pieces after 21 days. A subdermal pocket was created in each side of the dorsal midline of a rabbit, the fat pieces were grafted randomly into a pocket and the normal fat pieces into the other pocket as control. After 1, 3, 6, 9 and 12 months of implantation, the grafted fats were harvested, gross observation, weight measurement and histology were carried out. Number of the vessels stained with anti-CD34 antibody was counted out.

RESULTSVEGF concentrations in flaps were significantly higher (P < 0.05). The density of vessels in experimental groups increased significantly compared with that in control groups at 1 and 3 months, respectively (P < 0.01), and no significant differences in the survival rate of fat tissues between experimental and control groups were observed at 1 and 3 months (P > 0.05). The fat cells from the flaps survived after 12 months of fat plantation, while those in control groups disappeared after 6 months.

CONCLUSIONSThe survival rate and duration of grafted fat could be increased implanting the fat tissues from delayed fat flap, which may provide researchers with a new method for fat graft.

Adipocytes ; transplantation ; Adipose Tissue ; transplantation ; Animals ; Graft Survival ; Male ; Rabbits ; Surgical Flaps