OBJECTIVE:To optimize the extraction technique for Sidiming capsules.METHODS:The technical conditions for the water decoction and the alcohol precipitation were optimized respectively by the orthogonal experiment design L9(34)with hydrosoluble extract used as the index for the water decoction and the catalpol extract for alcohol precipitation.The content of Catalpol was determined by HPLC.RESULTS:The optimum conditions were as follows:decocting the crude drugs twice with 8-fold water,1 h each time.The physic liquor extracted by water was filtered,mixed and concentrated to 1.0 g?mL-1(crude drug),and then precipitated by 75% concentration of alcohol for 24 h.Then the physic liquor was filtered,concentrated and dried by microwave vacuum concentration dryer to obtain the dry ointment.CONCLUSION:The optimum extraction procedure is stable and reliable,and it can be used as the optimal extraction procedure of Sidiming capsules.

Objective To compare sleep electroencephalogram-modulated repetitive transcranial magnetic stimulation (SEM-rTMS) with routine repetitive transcranial magnetic stimulation (R-rTMS) in the treatment of primary insomnia. Methods One hundred and twenty six patients with insomnia were divided randomly into a SEM-rTMS group (44 cases) , a R-rTMS group (42 cases) and a sham rTMS group (40 cases). Each case was treated with rTMS for 30 min once a day for 10 d under double blinding of the patient and therapist. The clinical effect was observed along with Krakow sleep scores, electroencephalograms and blood pressure before and after a 10-day course of therapy and 30 days later. Results During rTMS treatment, some patients reported feeling sleepy, and this was confirmed by their EEGs at the time. After the entire course of therapy, 80% of the SEM-rTMS group and 45% of the R-rTMS group showed improvements, a significant difference. At the 30th day after therapy, the effect in the SEM-rTMS group was still significantly better than in the R-rTMS group. There were no obvious side effects in any group throughout the whole observation period. Conclusions SEM-rTMS is more effective and more reliable than R-rTMS. SEM-rTMS is safe to use with primary insomnia patients.

To explore the effect of high glucose on the NF-κB/IκB signal pathway in cultured rat glomerular mesangial cells. The results showed that high glucose increased the degradation of IκB-α and the translocation to nucleus of NF-κB. These changes could be reverted mostly by MG132, a proteasome inhibitor. It suggests that the activation of the NF-κB signal pathway by high glucose concentration may probably be via the ubiquitin-proteasome pathway.

Objective To evaluate the long-term efficacy and safety of donepezil in treating patients with Alzheimer's disease(AD).Methods Totally 86 patients with AD were randomly divided into control group(n =43)and treatment group(n =43).The control grou,p received conventional therapy with aniracetam,nimoldipine and ginkgo tablet,while the study group was administrated with donepezil(10 mg/d)on the basis of conventional therapy.The improvements of recognitive ability,mental state,activities of daily life were graded by mini-mental state examination (MMSE),Alzheimer's disease assessment scale-cog(ADAS-cog),activity of daily living(ADL)and global deterioration scale(GDS).The scores were compared between the groups before the treatment and 3,6,12,18,24,30,36,42,48,54,60,66 and 72 months after the treatment,respectively.Results The scores of MMSE,ADAS-cog and GDS after 3 months and ADL score after 6 months (t=2.361,-2.198,-1.790,-2.420,P＜0.05 or P＜0.01)were improved in treatment group than in control group with the best effects at 12 months(all P＜0.01)and the scores continued to decrease after 36 months.At 72 months,the score improvements in treatment group were 7.5 for MMSE,20.3 for ADAS-cog,19.5 for ADL,and 1.4 for GDS as compared with control group(all P ＜0.01).In contrast to pretreatment,there were statistically significant differences in the scores of MMSE,ADAS-cog and GDS at 3,6,12,18 and 24 months,and in the score of ADL at 6,12,18,24 and 30 months after treatment(P＜0.05 or P＜0.01).The differences in the scores of ADAS-cog and GDS after 24 months as well as MMSE and ADL after 30 months were not found(P＞0.05)between pre-treatment and post-treatment.Conclusions Donepezil might be long term effective and safe in slowing down the recognitive and overall function deterioration of AD.

AIM: To investigate the effects of protein C activator (PCA) from Agkistrondon acutus venom ( AAV) on the tension of thoracic aorta rings isolated from the rats with sepsis.METHODS:The model of sepsis was es-tablished by intraperitoneal injection of lipopolysaccharide ( LPS) .SD rats were randomly divided to 6 groups ( n=6 ):sham group, LPS group, PCA intervention group (LPS+PCA, PCA at doses of 0.1 mg/kg, 0.3 mg/kg and 0.6 mg/kg) and LPS+polymyxin B (at dose of 0.2 mg/kg) group.Using perfusion experiment in vitro, the tension of the aortic rings was measured by biological signal analytical system.RESULTS:The values of MABP, HR, LVDP and ±dp/dtmax were significantly lower in LPS group than those in sham group and LPS+PCA groups.Compared with sham group, the relaxa-tion response to acetylcholine ( ACh) and the contractile response of aorta rings induced by phenylephrine ( Phe) were sig-nificantly decreased in LPS group, which were increased significantly in PCA intervention group ( especially at dose of 0.6 mg/kg) compared with LPS group.The dose-response curve of aorta contraction with denuded endothelium induced by Phe shifted down significantly in LPS group compared with sham group, and no significant difference between LPS group and PCA intervention group was observed.Also no statistical difference was found in non-endothelium dependent relaxation of aortic rings induced by sodium nitroprusside among the groups.Pretreatment of N-nitro-L-arginine methl ester and methyl-ene blue increased the contraction amplitude of aortic rings induced by Phe.CONCLUSION:PCA from AAV effectively reverses the hypoergia of the vessels in rats with sepsis through protecting vascular endothelium, the mechanism of which may be mediated by inhibiting NO-GC-cGMP signal transduction pathway.

AIM:To investigate the effects of myeloid-related protein 8/14 ( MRP8/14 ) on the survival and apoptosis of human alveolar epithelial cell line A549, and to explore the role of nuclear factor-κB (NF-κB) in this process. METHODS:A549 cells were treated with different doses of MRP8/14 or stimulated with MRP8/14 for different time.The effect of MRP8/14 on the viability of A549 cells was determined by CCK-8 assay.The apoptotic rates were tested by flow cytometry.The nuclear translocation of NF-κB p65 was detected by Western blot and indirect immunofluorescence.Be-sides, the phosphorylation level of NF-κB p65 was determined by Western blot.NF-κB-specific inhibitor Bay 11-7082 was used to further analyze the role of NF-κB in the apoptosis induced by MRP8/14.RESULTS:The viability of the A549 cells was affected by MRP8/14 in a dose-and time-dependent manner.Flow cytometric analysis showed that the apoptotic rates were increased in the cells treated with MRP8/14 (P<0.01).In A549 cells administered with MRP8/14, NF-κB p65 was significantly phosphorylated and translocated into the nuclei, suggesting the activation of NF-κB signaling pathway. However, NF-κB-specific inhibitor Bay 11-7082 significantly attenuated the cell apoptosis induced by MRP8/14 ( P <0.01).CONCLUSION:NF-κB plays an important role in regulating the apoptosis of human alveolar epithelial cells in-duced by MRP8/14.

Objective To evaluate the clinical efficacy and safety of different full spectrum light times in treating patients with Alzheimer's disease (AD).Methods A total of 127 AD patients with sleep disorder were randomly divided into a blank group (n=34),a 30 min group (n=31),a 60 min group (n=33) and a 120 min group (n=29).After one month treatment by 10 000 lux full spectrum fluorescent light,the improvements of sleep quality,excessive daytime sleepiness,cognitive ability,mental state,dementia degree were graded by Pittsburgh sleep quality index (PQSI),Epworth sleepiness scale (ESS),Neuropsychiatric inventory (NPI),mini-mental state examination (MMSE),global deterioration scale (GDS).The scores were compared among the groups before the treatment and after the treatment respectively.Results (1) Compared with before treatment,the scores of PQSI,ESS,NPI of the 30 min group,60min group and 120min group were statistically significant (in 30 min group 14.4 ±5.2vs 11.7±4.9,14.4±4.1 vs 11.8±3.7,14.2±1.3 vs 10.9±1.7,t=2.071,2.609,8.446.P=0.043,0.011,0.000; in 60 min group13.4±4.0 vs 8.1±3.7,14.5±3.0 vs 9.4±2.0,13.7±5.8 vs 8.7±4.3,t=5.650,8.209,3.902,all P＜0.01 ;in 120 min group 14.0±3.2 vs 7.0±2.3,14.7-±2.3 vs 7.0± 1.9,14.9±3.6 vs 8.1±3.7,t=9.474,13.926,7.062,all P＜0.01),but the scores of MMSE,GDS were not statistical significances(all P＞0.05).(2)Compared with the blank group,the scores of PQSI,ESS,NPI of 30 min group,60 min group and 120 min group were statistically significant (30 min group t=1.936,4.524,2.482,P=0.031,0.000,0.016.60 min group t=5.945,5.153,7.319,all P=0.000.120 min group t=7.896,6.767,10.776,all P=0.000), but the scores of MMSE,GDS were not statistical significances(all P＞0.05).(3)Compared with the 30 min group,the scores of PQSI,ESS,NPI of 60 min group and 120 min group were statistically significances (60 min group t =3.288,2.694,3.354,P=0.002,0.009,0.001.120 min group t=4.615,3.930,6.303,all P =0.000),the scores of MMSE,GDS were not statistical significances (all P＞0.05).Compared with the 60 min group,the scores of ESS of 120 min group was statistically significant(t=4.854,P=0.000),but the scores of PQSI,NPI,MMSE,GDS were not statistical significances (all P ＞ 0.05).Conclusion It is demonstrated good curative effects that light therapy treat patients on AD patients in the matter of sleep quality,excessive daytime sleepiness,mental state,but have not apparent effect for their cognitive ability and dementia degree.And the effect of light therapy with 60 or 120 minutes is better than that of 30 minute,illumination time of 120 minutes is superior to that of 60 minutes in improving excessive daytime sleepiness.Light therapy has no obvious impacts in the cognitive ability and the degree of dementia in the patients with AD and has not appear obvious adverse reaction in the process of treatment.

AIM:To investigate the inhibitory effect of PDE4 inhibitor rolipram on the releases of inflammatory cytokines in mouse macrophages (RAW264.7 cells) induced by myeloid-related protein 8/14 (MRP8/14), and to explore the role of nuclear factor-κB (NF-κB) in this process.METHODS:RAW264.7 cells were treated with MRP8/14.The inflammatory cytokines TNF-α and IL-6 were determined by LiquiChip and qPCR.In contrast, RAW264.7 cells were pretreated with rolipram prior to MRP8/14 exposure.After MRP8/14 challenge, the inflammatory cytokines TNF-α and IL-6 were determined by LiquiChip and qPCR.Moreover, the phosphorylation level of NF-κB P65 was determined by Western blot.The nuclear translocation of NF-κB P65 in RAW264.7 cells was detected by indirect immunofluorescence.RESULTS:The results of LiquiChip and qPCR showed that MRP8/14 enhanced the expression of TNF-α and IL-6 (P<0.01),while pretreatment with rolipram markedly down-regulated the level of inflammatory cytokines induced by MRP8/14 (P<0.05).Meanwhile, MRP8/14 significantly activated the phosphorylation of NF-κB P65, and the protein expression of p65 in the nucleus was increased,while pretreatment with rolipram suppressed the phosphorylation of NF-κB P65 induced by MRP8/14.CONCLUSION:Rolipram may significantly reduce the releases of the inflammatory cytokines induced by MRP8/14 through the NF-κB signaling.

AIM:To explore the effects of stanniocalcin 2 (STC2) on the proliferation, migration and the process of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma HepG2 cells.METHODS:The expression levels of STC2 in the hepatocellular carcinoma cell lines and normal liver cells were assessed by Western blot.Colony formation assay was used to test the effect of STC2 on the proliferation of HepG2 cells.The effects of STC2 on the expression of proliferation-related molecules at mRNA and protein levels were determined by RT-qPCR and Western blot.The effect of STC2 on the migration ability was measured by Transwell assay.The mRNA and protein levels of vimentin and E-cadherin in STC2-overexpressing and-silencing cell lines were detected by RT-qPCR and Western blot.RESULTS:Compared with the normal liver cell line, the protein expression of STC2 was up-regulated in the hepatocellular carcinoma cell lines.The results of colony formation assay indicated that STC2 promoted the proliferation of HepG2 cells.STC2 significantly regulated the proliferation-related gene expression, such as cyclin D1.The results of Transwell assay showed that STC2 enhanced the migration ability of the HepG2 cells and influenced the EMT process.CONCLUSION:STC2 promotes the proliferation of HepG2 cells and affects the expression of proliferation-related genes.STC2 influences the process of EMT and promotes the migration of HepG2 cells.

AIM:To explore the mitochondrial pathway in the apoptosis of PC3 cells induced by PXD101 (also named as belinostat).METHODS:PC3 cells were treated with PXD101 at different doses or stimulated with PXD101 for different time.The effect of PXD101 on the viability of PC3 cells was measured by CCK-8 assay.The apoptotic rates and the mitochondrial membrane potential (MMP) were analyzed by flow cytometry.The protein levels of Bcl-2, Bax and cytochrome C (Cyt C) were determined by Western blot.The caspase-3 activity were tested by caspase-3 activity assay kit.RESULTS:The viability of the PC3 cells was inhibited by PXD101 in a dose-and time-dependent manner.Flow cytometric analysis showed that the apoptotic rates were increased in the cells treated with PXD101 (P<0.01).At the same time, PXD101 induced the decreases in MMP and Bcl-2, the release of Cyt C, and the increase in caspase-3 activity.CONCLUSION:PXD101 induces the apoptosis of human prostate cancer cell line PC3 by mitochondrial signal pathway.