Objective To construct an eukaryotic vector expressing short hairpin RNA(shRNA) of HIF-1α,and to observe its effects on location and metastasis of HeLa cells under hypoxic condition.Methods shRNA templates was developed based on HIF-1α gene sequence and then cloned into pSilencer2.1-U6-neo vector.The resultant plasmid was transfected into HeLa cells with Lipofectamine 2000.The cells were incubated in hypoxic condition.The HIF-1α protein and mRNA were detected by Western blot and RT-PCR.The colony formation assay and Transwell cabin assay were performed to measure the colony formation and metastasis.Results The plasmid pSilencer2.1-U6-neo-HIF-1α was successfully constructed and transfected into HeLa cells.The expression of HIF-1α in HeLa cells decreased,and the number of colony formation in soft agar and cells penetrating matrigel also decreased under hypoxic condition.Conclusion The shRNA expressing plasmid targeting at HIF-1α may suppress the location and metastasis of cervical carcinoma cells under hypoxic condition.

Objective To analyse clinical characteristics of patients with epilepsy induced by playing Mah-jong.Methods The medical records of 14 epileptic patients who were admitted at the Epilepsy Center of PUMC Hospital chief complaint of Mah-jong induced epileptic seizures from 1996 to 2000 were retrospectively reviewed.Results 12 patients showed generalized tonic-clonic seizures and 2 patients showed partial seizures with secondary generalization. Interictally, normal EEG were found in 13 patients, and one patient was discharged. None patients recept anticonvulsant therapy for preventing seizures. Conclusion Mah-jong induced epilepsy, a reflex epilepsy, is probably related to thinking,spatial task and decision making while playing mah-jong. It is benign, and could be easily prevented by stopping playing Mah-jong.

Objective To investigate the hemostatic effects and safety of the QuGouCao chloroform-water extract.Methods The hemostatic property of the extract was studied by applying the extract soaked gauze to the local wound surface of skin,liver and femoral artery in rabbits.LD50 test in Kuming mice was performed and the tissues of liver and kidney were collected for routine histochemical examination.The skin stimulus in rabbits and the sensitivity test in guinea pigs were conducted.Results The hemostatic time of wounds was obviously shorter(P

Objective To investigate the effect and mechanism of invigorating spleen,eliminating stasis, heat-clearing drugs(milk veteh,root of red rooted salvia,Chinese goldthread)on quality of ulcer healing (QOUH).Methods The rat models of chronic gastric ulcer was induced by acetic acid,the effects of QOUH of milk veteh,root of red rooted salvia and Chinese goldthread on the model of gastric Ulcer were observed, the expression of EGFR mRNA of mucosa were measured by in situ hybridization.Results Milk veteh,root of red rooted salvia and Chinese goldthread could improve the injury of gastric mucosa.Milk veteh,root of red rooted salvia increased the expression of EGFR mRNA in the tissue around peptic ulcer(PU). Conclusion Invigorating spleen,eliminating stasis,heat-clearing drugs can improve QOUH,it is one of possible mechanisms that the drugs increased the expression of EGFR mRNA in the tissue around PU.

Objective To investigate effect on telomerase activity of Hep-2 cells treated by anticancer drugs(hydroxycamptothecine, cisplatin and cytoxan).Methods By MTT method,we measured the 50% inhibitory concentration(IC 50 ) at 72h,and compared to untreated control cells. Telomerase activity of Hep-2 cells treated by the drugs in different concentration based on IC 50 for different time was observed by Telomeric Repeat Amplification Protocol with ELISA(TRAP-ELISA).Results Hydroxycamptothecine and cytoxan could inhibit proliferation of Hep-2 and down-regulate telomerase activity of Hep-2 cell. However, cisplatin promoted proliferation of Hep-2 and up-regulated telomerase activity of Hep-2 cell.Conclusions Hydroxycamptothecine and cytoxan could down-regulate telomerase activity of Hep-2 cell by direct or indirect pattern, which may correlate with drug concentration and time-dependent pattern.Cisplatin could up-regulate telomerase activity of Hep-2 cell, which mechanism is not clear.

Inhibitors of kinesin spindle protein (KSP) are a promising class of anticancer agents that cause mitotic arrest and induce apoptosis of tumor cells. A series of novel tetrahydro-beta-carboline derivatives were synthesized as kinesin spindle protein inhibitor and evaluated as potential antitumor agents. All compounds showed promising KSP inhibitiory activity. Compounds 8 and 9 exhibited better antitumor activity (Lung/A549, Stomach/AGS) than CK0106023 with GI50/IC50 values (1.07/1.62 and 1.46/3.27 micromol x L(-1), 1.09/>10 and 1.22/6.33 micromol x L(-1), respectively).

BACKGROUND: Tiantai I consists of gastrodia, Chinese angelica root, areca seed. It has been considered as the roles of invigorating the liver, nourishing marrow, heightening the intelligence and causing resuscitation. OBJECTIVE: To observe the effects of Tiantai Ⅰ on the abilities of learn ing and memory and the activity of central cholinergic system in mice withspontaneous Alzheimer disease. DESIGN: A randomized control observation. SETTING: Shenzhen Institute of Integrated Chinese and Western Medicine. MATERIALS: Male Kunming mice of 13 months old were raised to 21 months old, of which 52 with spontaneous Alzheimer disease werescreened. They were randomly divided into blank control group, westerndrug control group, Tiantai Ⅰ 6.80 and 20.41 g/kg groups, 13 mice in eachgroup. Another 13 aged mice with normal learning and memory abilitieswere selected as the normal control group at the same time. METHODS: Mice in the western drug control group were treated with0.6 mg/Kg Hydergine, those in the Tiantai Ⅰ 6.80 and 20.41 g/kg groupswere given intragastric administration of Tiantai Ⅰ of 6.80 and 20.41 g/kg, respectively, and those in the normal control group and blank control groupwere given double distilled water of the same volume. The learning andmemory results were examined by the step-down test. Freezing sections ofbrain tissue were prepared, acetylcholinesterase (AChE) fiber was showedaccording to the Hedreen method, and choline acetyltransferase (ChAT) was detected with Burt and Silver methods, the automatic image analysissystem for biomedical application was applied in the quantitative analysisof AChE fiber and ChAT activity. MAIN OUTCOME MEASURES: ① Effect of Tiantai Ⅰ on the abilities of learning and memory in mice with Alzheimer disease; ② AChE fiberarea density in temporal cortex and hippocampal CA1 region; ③ ChAT ac tivity in Meynert nuclei of basal forebrain. RESULTS: ① Tiantai Ⅰ in ameliorating the abilities of learning and mem ory in mice with spontaneous Alzheimer disease: The error times in theTiantai Ⅰ 6.80 and 20.41 g/kg groups, normal control group and westerndrug control group were obviously fewer than that in the blank controlgroup (5.08±2.18, 3.77±1.48, 3.25±1.27, 6.17±3.12, 9.31±3.65, P ＜ 0.05). ② AChE fiber area density: The AChE fiber area densities in temporalcortex were obviously higher in the Tiantai Ⅰ 6.80 and 20.41 g/kg groups, normal control group and western drug control group than in the blankcontrol group (20.19±4.18, 25.01±6.44, 26.38±6.52, 19.44±4.33, 14.36±3.21, P ＜ 0.05). The AChE fiber area densities in hippocampal CA1 region were obviously higher in the Tiantai Ⅰ 20.41 g/kg groups and nor mal control group than in the blank control group, western drug controlgroup and Tiantai Ⅰ 6.80 g/kg group (13.88±3.07, 14.11±4.36, 6.57±2.30, 10.13±2.94, 9.63±2.40, P ＜ 0.05). ③ ChAT activity: It was obviously high er in the TiantaiI 20.41 and 6.80 g/kg groups, western drug control groupand normal control group than in the blank control group (0.683±0.179, 0.589±0.179, 0.552±0.173, 0.705±0.141, 0.354±0.131, P ＜ 0.05). CONCLUSION: Tiantai Ⅰ can significantly improve the disturbances oflearning and memory in mice with spontaneous Alzheimer disease, and itcan increase the activity of central cholinergic system in a dose-dependentmanner.

BACKGROUND: The early and medium medication has been proved to have a certain effect to patients with Alzheimer disease, can delay its development.OBJECTIVE: To investigate the effect of No. 1 recipe on learning and memory function of spontaneous Alzheimer disease models.DESIGN: A randomized and controlled trial.SETTING: Shenzhen City Institute of Integration of Traditional Chinese and Western Medicine.MATERIALS: The experiment was completed at the grade Ⅱ Laboratory of Shenzhen City Institute of Integration of Traditional Chinese and Western Medicine. The experimental animals were mice of Kunming strain.METHODS: Totally 52 Kunming mice, aged 21 months, with spontaneous Alzheimer disease (memory impairment) were chosen, and at random divided into 4 groups: senile dementia, western drug control, tiantai No. 1recipe 6.80 g/kg and 20.41 g/kg groups; besides 13 mice with normal learning and memory power were chosen as normal senile group. Hydergine 0.6 mg/kg was given for mice in western drug control group, tiantai No. 1recipe (orthogonally optimized) 6.80 g/kg and 20.41 g/kg were respectively given for mice in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups.The medicines above were all prepared into 0.5 mL solution and given by garage, for consecutive 60 days. The mice in normal senile and senile dementia groups were given double distilled water of the same volume. The learning memory power was tested by jumping stand, comparing the numbers of learning error and memory error, latent escaping periods of learning training and latent safe platform of memory test.MAIN OUTCOME MEASURES: The improving effect of tiantai No. 1recipe on learning and memory function of spontaneous Alzheimer disease mice.RESULTS: All animals involved in the trial entered the final result analysis without any loss. The error numbers of learning and memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were smaller than that in senile dementia group (5.08±2.18, 3.77±1.48, 9.31±3.65; 1.85±1.21, 1.54±0.88,4.27±2.58), and the error number of learning in tiantai No. 1 recipe 6.80 g/kg group was more than that in tiantai No. 1 recipe 20.41 g/kg group. The escape latent period of learning training was smaller than that in the senile dementia group and the safe platform latent period of memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were longer than that in senile dementia group [(18.9±7.5), (19.9±5.9), (33.7±9.3); (91.7±32.0),(101.5±40.9), (43.6±20.7)s], and the safe platform latent period of memory in tiantai No. 1 recipe 6.80 g/kg group was smaller than that in 20.41 g/kg group.CONCLUSION: There was obvious intellectual disturbance in spontaneous Alzheimer disease mice, tiantai No. 1 recipe can remarkably improve their learning and memory impairments, with a relationship of dose-effect.

BACKGROUND:Neuron nitrogen monoxide(NO) is related to synaptic plasticity and is the key transmitter of normal learning and memory.Neuronal nitric oxide synthase(nNOS) is the biological synthetic enzyme of NO in neurons.The reports have been fewer yet on cerebral nNOS activity in Alzheimer disease(AD) patients,especially on changes in positive neural fibers of nNOS in temporal lobe cortex and hippocapmus CA1 region. OBJECTIVE:To observe cerebral nNOS activity in AD patients, especially on changes in positive neural fibers of nNOS in temporal lobe cortex and hippocapmus CA1 region and effects of Tiantai Ⅰ on nNOS activity. DESIGN:A randomized and controlled trial. SETTING:Institute of Collaboration of Chinese and Western Medicine, Shenzhen City. MATERIALS:The experiment was performed in No.2 Grade Animal Experimental Room of Institute of Collaboration of Chinese and Western Medicine,Shenzhen City.A total of 65 Kunming mice were employed in the experiment. METHODS:The experimental animals were bred till 21 months old and the mice with senile dementia were screened from the aged ones according to the international general standards and methods.The experimental animals were randomized into 4 groups,named as senile dementia group,western drug control,group with small dosage of Tiantai Ⅰ and group with large dosage of Tiantai Ⅰ .In addition,a group with normal learning and memory of the senile mice(simply called senile normal group) was designed.There were 13 mice in each group.In western drug control,hydergine 0.6 mg/kg was prescribed; in the groups of small and large dosage of Tiantai Ⅰ ,the dosages were 6.80 g/kg and 20.41 g/kg respectively,continuously for 60 days.In senile normal group and senile dementia group,the bi distilled water of equal dosage was applied for perfusion.The scores of learning and memory were determined by step down test. Slices of brain tissues were prepared with frozen,nNOS activity was displayed by NBT histochemistry method and the analysis was done systematically with auto micrography in quantity. RESULTS:Learning and memory were significantly decreased in mice with senile dementia(P< 0.01),the positive reaction of nNOS was remarkably weaker compared with normal aged mice.It was the first time to discover that the surface densities of nNOS positive neural fibers in temporal lobe cortex and hippocampus CA1 region [(9.48± 2.09),(5.11± 2.74) respectively]were remarkably lower than those of the normal aged mice [(17.75± 3.47),(11.95± 2.00) respectively](P< 0.01). Simultaneously, it was also the first time to discover that normal correlation was apparent between nNOS activity of cortex and hippocampus and the scores of learning and memory.Tiantai Ⅰ could significantly increase the surface densities of nNOS positive neural fibers in cortex and hippocampus in senile dementia mice, of which, the values of the small dosage were (11.57± 2.44) and (8.18± 2.92) respectively and of the large dosage were (12.59± 2.88) and (9.33± 2.18) respectively,(P< 0.01- 0.05). CONCLUSION:The hypo activity of central nNOS in senile dementia mice resulted in significantly decreased positive neural fibers of nNOS in cerebral cortex and hippocampus.The surface densities of nNOS positive neural fibers in temporal lobe cortex and hippocampus CA1 region were significantly related to the scores of learning and memory.Tiantai Ⅰ could remarkably improve the disturbance of learning and memory in spontaneous senile dementia model and enhance its nNOS activity in cerebral cortex and hippocampus.

Objective To analyze the composing experience of Menghe physician Ma Peizhi for cough by TCM inheritance support system. Methods The prescriptions for cough of Ma Peizhi were collected, frequency and association of drugs were analyzed by using data mining methods such as revised mutual information, complex system entropy cluster and unsupervised hierarchical cluster. Results Based on the analysis of 57 prescriptions, the frequency of each herb and association rules among the herbs were computed, 18 core combinations and 9 new prescriptions were mined from the database. Conclusion Menghe physicians Ma Peizhi is well experienced in expelling wind and opening the inhibited lung, dissolving phlegm and relieving cough. TCM inheritance support system can be used to analyze clinical experience of old TCM doctor.