OBJECTIVE To investigate the intervention effect and its potential mechanism of Yifei xuanfei jiangzhuo formula by inhibiting mitochondrial fission in a vascular dementia （VaD） model rats. METHODS VaD rat model was established by bilateral common carotid artery ligation. The experimental animals were randomly divided into sham operation group （SHAM）， model group （MOD），Yifei xuanfei jiangzhuo formula low-dose group （YFXF-L）， Yifei xuanfei jiangzhuo formula high-dose group （YFXF-H）， and Donepezil hydrochloride group （positive control）， with 9 animals in each group. After 30 days of intervention， the spatial learning memory ability was assessed by Morris water maze experiment； HE staining was used to observe histopathological changes in CA1 area of hippocampus； ELISA was used to detect the levels of serum inflammatory factors [interleukin-1β （IL-1β） and IL-4]； Western blot was used to detect the expressions of heat shock protein 90 （HSP90）/mixed lineage kinase domain-like protein （MLKL）/dynamin-related protein 1 （Drp1） pathway-related proteins， mitochondrial fusion proteins （MFN1， MFN2）， and adenosine triphosphate synthase 5A （ATP5A） in hippocampal tissues. The immunohistochemistry was used to detect the level of phosphorylated MLKL （p-MLKL）； real-time fluorescence quantitative PCR was adopted to detect mRNA expressions ofHSP90， MFN1， MFN2 and ATP5A. RESULTS Compared with SHAM group， the escape latency of rats in the MOD group was significantly prolonged， the number of crossing the platform was significantly reduced， and the hippocampal tissues showed typical neuronal damage characteristics， the positive expression level of p-MLKL and the serum level of IL-1β significantly increased， while the serum level of IL-4 significantly decreased， the protein and mRNA expression of HSP90， as well as the protein expressions of p-MLKL/MLKL and p-Drp1（Ser616）/Drp1 were all significantly increased in hippocampal tissue， the protein and mRNA expressions of MFN1， MFN2 and ATP5A， and protein expression of p-Drp1（Ser637）/Drp1 were all significantly decreased （P＜0.05）. After the intervention of Yifei xuanfei jiangzhuo formula， above indicators in each treatment group were all significantly reversed （P＜0.05）. CONCLUSIONS Yifei xuanfei jiangzhuo formula may alleviate neuronal damage and neuroinflammatory responses in VaD rats by regulating the HSP90/MLKL/Drp1 signaling pathway， inhibiting mitochondrial fission， thereby maintaining mitochondrial dynamic balance and improving mitochondrial function.

Objective To construct a RNA interference lentiviral vector aimed at human ARK5 (AMPK-related protein kinase 5) gene and explore its effect on the biologic behavior of human gastric cancer SGC7901 cells.Metho ds Targeting human ARK5 mRNA coding sequence, we designed three specific short hairpin RNAs (shRNAs) and constructed the lentiviral vector,then infected human gastric cancer SGC7901 cells with this vector.Afterwards, we used qPCR and Western blot for detecting the silencing effect on ARK5 gene,MTT colorimetric assay to measure the cell proliferation , cell scratch test for cell migration and Transwell for cell invasion, and flow cytometry analysis for apoptosis in cells treated with glucose starvation and TNF-α.Results Sequencing proved that the recombinant lentiviral vector containing ARK5-shRNA-3 was constructed successfully.Real time fluorescent quantitative PCR assay showed that the expression abundance of ARK5 gene in the normal control group, negative control group and ARK5-shRNA-3 infected group were 1.002+0.082, 1.001+0.050 and 0.140+0.003, respectively, showing a statistically significant difference (P<0 .01).Cell scratch test showed that the cell migration rate of ARK5-shRNA-3 infected group was (38.5+4.3 )%, significantly lower than that of the normal control group [(72.4+6.4)%] and negative control group [(75.1+7.1)%, P<0.01].The results of Transwell test showed that the number of penetrating cells in the normal control group, negative control group and ARK5-shRNA-3 transfection group were 257.4±12.3, 245.7±11.6, 112.5±7.8, with a significant difference (P<0.01).After glucose starvation and TNF-α-treatment for 24 h, the cell death rate of the normal control group, negative control group and ARK5-shRNA-3 group were (11.7±3.2)%, (12.3±2.6)% and (30.8±4.3)%, respectively, showing that the cell apoptosis rate of ARK5-shRNA-3 transfected group was significantly higher than that of the normal control and negative control groups (P<0.01).Conclusions We have successfully constructed a recombinant lentiviral vector which can efficiently silence ARK5 gene.Using it we can inhibit the proliferation, migration, invasion of tumor cells, and promote cell apoptosis under the condition of TNF-αtreatment and glucose starvation.

Objective To construct a RNA interference lentiviral vector aimed at human ARK5 (AMPK-related protein kinase 5) gene and explore its effect on the biologic behavior of human gastric cancer SGC7901 cells.Metho ds Targeting human ARK5 mRNA coding sequence, we designed three specific short hairpin RNAs (shRNAs) and constructed the lentiviral vector,then infected human gastric cancer SGC7901 cells with this vector.Afterwards, we used qPCR and Western blot for detecting the silencing effect on ARK5 gene,MTT colorimetric assay to measure the cell proliferation , cell scratch test for cell migration and Transwell for cell invasion, and flow cytometry analysis for apoptosis in cells treated with glucose starvation and TNF-α.Results Sequencing proved that the recombinant lentiviral vector containing ARK5-shRNA-3 was constructed successfully.Real time fluorescent quantitative PCR assay showed that the expression abundance of ARK5 gene in the normal control group, negative control group and ARK5-shRNA-3 infected group were 1.002+0.082, 1.001+0.050 and 0.140+0.003, respectively, showing a statistically significant difference (P<0 .01).Cell scratch test showed that the cell migration rate of ARK5-shRNA-3 infected group was (38.5+4.3 )%, significantly lower than that of the normal control group [(72.4+6.4)%] and negative control group [(75.1+7.1)%, P<0.01].The results of Transwell test showed that the number of penetrating cells in the normal control group, negative control group and ARK5-shRNA-3 transfection group were 257.4±12.3, 245.7±11.6, 112.5±7.8, with a significant difference (P<0.01).After glucose starvation and TNF-α-treatment for 24 h, the cell death rate of the normal control group, negative control group and ARK5-shRNA-3 group were (11.7±3.2)%, (12.3±2.6)% and (30.8±4.3)%, respectively, showing that the cell apoptosis rate of ARK5-shRNA-3 transfected group was significantly higher than that of the normal control and negative control groups (P<0.01).Conclusions We have successfully constructed a recombinant lentiviral vector which can efficiently silence ARK5 gene.Using it we can inhibit the proliferation, migration, invasion of tumor cells, and promote cell apoptosis under the condition of TNF-αtreatment and glucose starvation.

Objective To study the relationship between expression of matrix metalloproteinases-7 (MMP-7) and clinicopathological characteristics in patients with primary non-smaU cell lung cancer(NSCLC). Methods MMP-7 in 20 normal people and 60 advanced NSCLC patiens were detected with reverse-transcription-polymerase-chain-reaction. Gelatum image analysator analyzed the result. Results The amount of MMP-7 was less in normal people (30.000) than in NSCLC patients(41.231) significantly(P<0.05); the level of MMP-7 was no correlated with gender, age, pathology pattern, tumor size, was inverse correlation with differentiation, and was positive correlation with clinical stages(P <0.05). Conclusion The level of MMP-7 is closely correlated with tissue differentiation and clinical stages of NSCLC, which may serve as a parameter for determining tumor invasion and metastatic.

BACKGROUNDTo evaluate the efficacy and toxicity of ifosfamide and cisplatin in the treatment of advanced non-small cell lung cancer.

METHODSFifty-six patients with advanced non-small cell lung cancer were treated by combination chemotherapy of ifosfamide and cisplatin for two cycles: ifosfamide 1.5-2.0g/m² iv drip on day 1-4, mesna 400mg iv at 0,4,8 hours after using ifosfamide; DDP 25-30mg/m² iv drip on day 5-7. The response, toxicity, relievable period and survival period were evaluated.

RESULTSThe total response rate was 50.0%. The response rate of patients in primary treatment was 52.8% and that of return cases was 45.0% (P > 0.05). The median relievable period was five months. The median duration of survival (MDS) was nine months. The major toxicity was inhibition of bone marrow, especially of leukocyte and platelet.

CONCLUSIONSCombination chemotherapy of ifosfamide and cisplatin is effective in the treatment of advanced non-small cell lung cancer including the return cases, and the toxicity is tolerable. If G-CSF is used as a complementary therapy, this regimen could be quite clinically valuable.

The effect of plasma parathyroid hypertensive factor (PHF)from 153 essential hypertensive subjects on pressor response in normotensive rats was evaluated and plasma levels of parathyroid hormone (PTH) in these patients were measured. The results showed that PHF from 61 patients could induce a delayed increase in blood pressure in the normotensive rats. Mean arterial pressure (MAP) began to increase at 10 minutes after injection and a peak response was reached at 50 minutes while pre-injection MAP level was recovered at 70 minutes. On the other hand, plasma PHF in remaining 92 patients failed to induce a pressor effect. Plasma PTH in the 153 patients were considerably higher than those of the control, and the plasma PTH from the two groups, either with or without PHF, showed significant difference in comparison with the control, but plasma PTH between these two groups did not show significant difference. The results have suggested that pressor effect was independent of PTH but depends on PHF.