Objective: To investigate the induction effect of oxymatrine on the apoptosis process in rat hepatic stellate cell line HSC-T6 and to define its impact on telomerase activity and mRNA expression of subunit telomerase reverse transcriptase (rTERT). Methods: HSC-T6 cells were cultivated with different concentration of oxymatrine for different time periods. Effect of oxymatrine on the growth inhibition of HSC-T6 cells was analyzed by MTT assay. Apoptosis of HSC-T6 cells was detected by flow cytometry. Telomerase activity was determined by TRAP-PAGE-silver staining and the expression of rTERT mRNA was examined by RT-PCR assay. Results: Oxymatrine significantly suppressed the growth of HSC-T6 cells and induced apoptosis, and also reduced the activity of telomerase and inhibited the rTERT-mRNA expression in HSC-T6 cells. Conclusion: The function that oxymatrine inhibits the proliferation of HSC-T6 cells may be associated with its action on the cellular telomerase and telomerase rTERT-mRNA activity.

Objective: To explore the factors affecting energy efficiency factor (EEF) in high intensity focused ultrasound for adenomyosis. Methods: Data of 130 patients with adenomyosis treated with high intensity focused ultrasound were retrospectively analyzed. The factors affecting EEF in high intensity focused ultrasound were identified with difference and correlation analyses. And a multiple linear regression equation was obtained by multivariate analysis. Results: The course of disease (X1), abdominal wall scar (X2), gonadotropin releasing hormone agonist (GnRH-a) pretreatment (X3), blood flow grade (X4), abdominal wall thickness (X5) and target-skin distance (X6) were the influencing factors of EEF in high intensity focused ultrasound with the multiple linear regression equation as Y=-17.742+1.153X1+14.927X2-13.846X3+4.713X4+1.422X5+0.227X6. Conclusion: High intensity focused ultrasound treatment presents lower EEF and higher ablation efficiency in patients with adenomyosis who have short course of disease, no abdominal wall scar, GnRH-a pretreatment, little blood flow in the lesion, thin abdominal wall and short target-skin distance.

OBJECTIVETo study the effect of melatonin on the expressions of glial fibrillary acidic protein (GFAP), nuclear factor-κB (NF-κB p65) and synaptophysin in mice of different ages.

METHODSTwenty young male B6C3F1 mice (5.5 months) and 20 aged mice (26 months) were both divided into control and melatonin treatment (daily dose of 0.04 mg/kg) groups. After 2.5 months of treatment, the brain tissues of the mice were collected to examine the expressions of GFAP, NF-κB and SYN by immunohistochemistry.

RESULTSIn the control groups, the expression of NF-κB p65 in the brain tissue increased with age, whereas a reverse change was found in melatonin-treated aged rats (P<0.05). Synaptophysin expression also decreased with age, but melatonin treatment significantly enhanced its expression in aged mice (P<0.05). GFAP expression in the brain tissue increased with age regardless of melatonin treatment (P>0.05).

CONCLUSIONGFAP expression is almost not affected by melatonin treatment in aged mice. Melatonin can reduce the expression levels of NF-κB p65 and synaptophysin in the brain tissue to protect the brain and slow down the aging process.

Aging ; metabolism ; Animals ; Brain ; metabolism ; Chimera ; Glial Fibrillary Acidic Protein ; Male ; Melatonin ; pharmacology ; Mice ; Mice, Inbred C3H ; Mice, Inbred C57BL ; NF-kappa B ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Synaptophysin ; genetics ; metabolism ; Transcription Factor RelA ; genetics ; metabolism

Objective To reduce the animal component contamination for human embryonic stem cells ( hESCs ) and to simplify hESCs culture process , we develop a new coating substrate which can support the hESCs growth without dif -ferentiation, and is easy to store and use. Methods Mouse embryonic fibroblasts(MEF)were fixed on the surface of plate by methanol.hESCs were cultured on this new substrate and were passaged every 5 to 6 days.After 10 passages, we checked the cell morphology , alkaline phosphatase expression , embryonic specific markers and the differentiation ability in vitro.Results After 10 passages , the hESCs grew well on this new substrate and maintained the typical hESCs morpholo -gy.Alkaline phosphatase staining was positive .Immunofluorescence staining showed that the expressions of Oct 4, SSEA4, Tra-1-60 were positive .The cells formed embryoid body in vitro .Conclusions This methanol-fixed MEF substrate can support the growth of undifferentiated hESCs .The coating material can be produced in large scale and stored for a long time.It provides a new and relatively easy way to amplify hESCs .

OBJECTIVETo study the effects of Ginkgo biloba extract (GBE) on the pharmacokinetics and pharmacodynamics of warfarin and observe the anticoagulant activity of GBE.

METHODA randomized, double-blinded, placebo-controlled, two-way cross-over trial was conducted. Twelve healthy volunteers (sex ratio was 1: 1) were randomized into two groups and received GBE (three pill, tid) or placebo (three pill, tid) for 5 weeks respectively. the subjects received a single dose of warfarin (5 mg) on the day 29. Blood samples for pharmacokinetics and pharmacodynamics assessment were collectd.

RESULTCompared with placebo, BE had no significant pharmacodynamics effects on warfarin and had no effects on prothrombin time (PT) and activated partial thromboplastin time (APTT). GBE extract increased C(max), AUC(0-144 min), AUC(0-infinity), t1/2, of warfarin significantly and decreased CL(F) of warfarin significantly (P < 0.05), and there were no singnificant difference of V(d) (F).

CONCLUSIONGBE has limited effects on the pharmacokinetics but no effects on the pharmacodynamics of single dose warfain in health subjects. GBE has no effects on clotting process alone.

Adult ; Anticoagulants ; pharmacology ; Double-Blind Method ; Female ; Ginkgo biloba ; Herb-Drug Interactions ; Humans ; Male ; Plant Extracts ; pharmacology ; Warfarin ; blood ; pharmacology ; Young Adult

OBJECTIVETo investigate the mechanisms of Buyang Huanwu Decoction (BYHWD) by observing its effects on expressions of angiopoietin-1 (Ang-1) and the endothelial-specific receptor tyrosine kinase (Tie-2) mRNA in damaged region of rats' brain after intracerebral hemorrhage (ICH).

METHODSOne hundred and sixty Sprague-Dawley rats were randomly divided into four groups, 10 in the normal control group, 60 in the sham-operative group, 60 in the ICH model group, and 30 in the BYHWD-treated group. The ICH model was established by injecting collagenase type VII 0.5 U stereotaxically into right globus pallidus. Animals in the BYHWD-treated group were administered orally with BYHWD, while animals in the sham-operative group and the ICH model group were administered orally with equal volume distilled water, and those in the normal control group drank water freely. The positional variations of the expression of Ang-1 and Tie-2 in the sham-operative group and the model group were assayed by immunohistochemistry on dayl, 4, 7, 14, 21 and 28 after modeling, in the meantime, the dynamic changes of Ang-1 and Tie-2 mRNA expressions in all groups were assayed by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSNo significant expression of Ang-1 and Tie-2 in brain of rats in the normal or the sham-operative group was found during the experiment. In the model group, the Ang-1 and Tie-2 positive micrangio-segments appeared at the edge of clot on day 1 to day 4, they gradually penetrated to hematoma area from day 7; with Ang-1 and Tie-2 mRNA expressed from day 1, but very weak until day 4, showing no significant difference to that on day 1; thereafter, they increased gradually, and reached the peak on day 28 (P <0.05). While the two expressions in the BYHWD treated group reached the peak on day 21, and from day 7 to day 28, they were all significantly higher than those in ICH model group at the corresponding time points (P <0.01).

CONCLUSIONBYHWD can promote the up-regulation of Ang-1 and Tie-2mRNA expressions in brain of intracerebral hemorrhagic rats, which might accelerate the angiogenesis in the reconstruction of microvascular network in the damaged zone, and thus facilitating the repairing of damaged tissue.

Angiopoietin-1 ; genetics ; metabolism ; Animals ; Brain ; drug effects ; metabolism ; Cerebral Hemorrhage ; drug therapy ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Receptor, TIE-2 ; genetics ; metabolism

Objective: To investigate the effect of endocrine gland-derived vascular endothelial growth factor (EG-VEGF) on the malignant behavior of colon cancer LoVo cells. Methods: The LoVo cells treated with different concentrations (50, 100, and 200 μg · L-1) of EG-VEGF were used as EG-VEGF groups, and the colon cancer LoVo cells cultured with solution without EG-VEGF were regarded as control group. The proliferation activity of LoVo cells was determined by MTT method; flow cytometry (FCM) was used to detect the percentages of LoVo cells at different cell cycles in various groups; Wound scratch assay test and Transwell cell migration assay were used to detect the migration rates of LoVo cells and the number of migration cells. Results: The MTT results showed that compared with control group, the proliferation activities of LoVo cells in 50, 100, and 200 μg · L-1 EG-VEGF groups were increased significantly (P<0. 05); with the increasing of concentration of EG-VEGF, the proliferation activity was increased apparently. The FCM results showed that compared with control group, the percentage of colon cancer LoVo cells at G0/Gi phase in 100 μg · L-1 EG-VEGF group was decreased (P<0. 05) and the percentage of colon cancer LoVo cells at S phase was increased (P<0. 05), but the percentage of colon cancer LoVo cells at G2 + M phase did not change significantly. The wound scratch assay results showed that compared with control group, the migration rate of LoVo cells in 100 μg · L-1EG-VEGF group was increased (P< 0. 05). The Transwell assay results showed that compared with control group, the number of migration cells in 100 μg · L-1 EG-VEGF group was increased significantly (P<0. 05). Conclusion: EG-VEGF can obviously promote the proliferation and migration of colon cancer LoVo cells, and EG-VEGF may be associated with the malignant development of colon cancer.

Objective: To evaluate the capability of magnetic resonance imaging 3D-sampling perfection with application optimize contrasts using different flip angle evolution (MRI 3D-SPACE) full-volume scanning sequence to detect lower extremity deep vein thrombosis (LEDVT), and to explore the characteristics of the images of LEDVT and surrounding soft tissue at different stages. Methods: MRI 3D-SPACE full volume T1WI and T2WI scanning of lower limb were performed on 45 patients diagnosed of LEDVT via digital subtraction angiography (DSA) by Siemens Prisma 3T MRI scanner. 43 cases completed scanning. The location and quantity of vascular segment lesions were recorded, of which the diagnostic sensitivity, specificity and accuracy were calculated comparing to the gold standard of DSA results. The characteristics of magnetic resonance imaging signals of thrombus and thrombus surrounding soft tissue at different times were summarized. Results: According to the location of thrombosis, there were 15 cases of central type, 17 cases of peripheral type and 11 cases of mixed type. In all imaging levels, 272 segments of embolic vessels were found in 3D-space T1WI and 261 segments in 3D-space T2WI. Compared with the DSA results, the sensitivity, specificity, and accuracy of the two groups in the diagnosis of deep venous thrombosis were: T1 WI: Common iliac vein 97.10%, 97.90%, 98.20%; External iliac vein 96.80%, 97.60%, 97.10%; Internal iliac vein 96.40%, 97.20%, 97.20%; Femoral vein 98.10%, 98.20%, 98.50%; Popliteal vein 94.40%, 93.50%, 93.70%; The anterior tibial vein is 94.30%, 88.50% and 90.40%. The posterior tibial vein is 93.40%, 85.20%, 85.30%. T2WI: Common iliac vein 97.10%, 97.90%, 97.10%; External iliac vein 96.50%, 96.20%, 96.00%; Internal iliac vein 96.20%, 96.60%, 96.20%; Femoral vein 98.10%, 98.20%, 98.50%; Popliteal vein 93.40%, 92.50%, 92.70%; The anterior tibial vein is 90.30%, 86.50% and 85.40%; Posterior tibial vein 89.40%, 84.20%, 83.30%. According to the onset time, 17 cases in the acute phase, the thrombus was equal or slightly higher and uneven signal in the two groups of sequences, the perivascular and muscle tissue gap signal increased; 21 cases in the subacute phase, the thrombus was high in both groups signal; 5 cases in the chronic phase, the thrombus showed equal or slightly lower signal in the two groups of sequences, the blood vessel wall was irregular, and the surrounding soft tissue was mild or no obvious edema. Conclusions MRI 3D-space sequence full volume scanning technology has high sensitivity, high specificity and high accuracy for the diagnosis of lower extremity deep vein thrombosis, and can reflect the characteristics of thrombus and soft tissue signal around thrombus in different periods. It has high application value for the early detection of thrombosis and the stage diagnosis of the course of disease.

OBJECTIVE: To investigate the effect of oral administration of insulin on insulitis beta cell apoptosis and diabetes in non-obese diabetic (NOD) mice, and to explore the mechanism of immune tolerance induced by insulin. METHODS: Eighty-six female NOD mice were randomly divided into an insulin group (n=43) and a phosphate buffered saline (PBS) group (n=43). From 4 weeks of age, the recombinant human insulin (Humulin R) 1 mg (70 microL) was administrated in the oral insulin group and 70 microL PBS in the control group respectively, twice per week before 12 weeks of age and then once weekly until 30 weeks. Insulitis and beta cell apoptosis of islets were observed at 12 weeks. IL-4 and IFN-gamma in the sera were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of I-Abeta(g7), IL-4, IFN-gamma, IL-1beta, Fas and TGF-beta mRNA of islets, and IL-4, IFN-gamma, TGF-beta mRNA of Peyer's patch were measured by reverse transcription-polymerase chain reaction (RT-PCR) at 12 weeks. RESULTS: The incidences in the insulin group were significantly lower than those in the PBS group (55.6% vs 85.7% at 30 weeks, 70.4% vs 96.4% at 52 weeks, P<0.05). The insulitis scores in the insulin group were lower than those in the PBS group, but there was no statistical significance. Fas expression on islets and apoptotic beta cell rates in the insulin group were lower than those in the PBS group (P<0.05). In the insulin group, serum IL-4 levels were higher, and IFN-gamma levels were lower than those in the PBS group (P<0.05). The levels of I-Abeta(g7), IFN-gamma, IL-1beta and Fas mRNA transcription in islets and IFN-gamma mRNA transcription in Peyer's patch were both lower in the insulin group, and IL-4, TGF-beta mRNA levels were higher than those in the PBS group (P<0.05). CONCLUSION The specific autoantigen insulin may induce the immune tolerance and prevent the diabetes in NOD mice, but it cannot block the progression of insulitis. Oral administration of insulin can induce the regulatory T cells, and make Th1 to Th2 cytokine shifts in the system and islets, thus preventing the Fas-mediated beta-cell apoptosis and diabetes.
Administration, Oral ; Animals ; Apoptosis ; drug effects ; Cytokines ; metabolism ; Diabetes Mellitus, Type 1 ; drug therapy ; pathology ; Female ; Insulin, Regular, Human ; administration & dosage ; pharmacology ; Islets of Langerhans ; cytology ; drug effects ; Mice ; Mice, Inbred NOD ; Th1-Th2 Balance ; fas Receptor ; metabolism

OBJECTIVE: To investigate the effect of human papillomavirus types 16E6 on the sensitivity of chemotherapy for cervical carcinoma in different p53 genotype cell lines. METHODS: The apoptosis rates of each group were detected by AO/EB, immunofluorescence and Annexin V/PI stained methods. The expressions of protein HPV16E6 and p53(mt) after the treatments of different concentration of DDP were detected by Western blot. HPV16E6 mRNA in C33A, C33A-E6, C33A-P, and CaSki cell lines under different DDP treatments were detected by RT-PCR. RESULTS: AO/EB and Annexin V/PI stained tests showed that the apoptosis rates of C33A, C33A-E6, C33A-P, and CaSki cells were significantly increased when DDP concentration increased. Western blot showed that the HPV16E6 protein could be detected only in C33A-E6 and CaSki cell lines. The expression of HPV16E6 protein in C33A-E6 and CaSki cell lines gradually decreased and was hardly detected with increased dosage of DDP and the prolonged treatment time (P<0.01), and slightly increased in C33A-E6 and Caski cell lines without the treatment, but there was no significant difference between them (P>0.05). Protein p53(mt) persistently expressed in C33A-E6, C33A, and C33A-P cell lines following the increased dosage of DDP and the prolonged treatment time(P>0.05), while it couldn't be found in CaSki cell line. RT-PCR showed that without DDP intervention, there was no significant difference of HPV16E6 mRNA in C33A-E6 and CaSki cell lines within 24 h.The HPV16E6 mRNA in C33A-E6 cell line expressed much higher than that in CaSki (P<0.05), and HPV16E6 mRNA of 2 cell lines expressed much higher at 48 h than at 24 h (P<0.05).The expression of HPV16E6 mRNA in C33A-E6 and CaSki cell lines gradually decreased with the increased DDP and prolonged treatment time (P<0.01), while there was no significant difference between C33A-E6 and CaSki cell lines under the same DDP concentration (P>0.05). CONCLUSION Effect of HPV16E6 on the sensitivity of chemotherapy for cervical carcinoma cell lines is not markedly related with the different p53 genotype forms(p53(mt)/p53wt ). HPV16E6 may affect the proliferation and sensitivity of chemotherapy in C33A cell line through other mechanism.
Antineoplastic Agents ; pharmacology ; Apoptosis ; genetics ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Female ; Genotype ; Human papillomavirus 16 ; genetics ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Repressor Proteins ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; Uterine Cervical Neoplasms ; pathology ; virology