Objective To explore the effects of Kangshuaiyizhi capsule on ChAT/AchE and NE, DA and 5-HT levels in the brain tissue of aging model rats, and explore its effect of protecting cerebral function. Methods Totally 72 rats were randomly divided into normal group and model group. The subacutely aging model rats were made by injecting D-gal (0.125 g/kg) into abdominal cavity continually, then aging rats were divided by random number table into model group, Naofukang group and Kangshuaiyizhi high-, low-dose group. After intervented with correspongding drugs for 60 days, activity of ChAT and AchE, cerebral cortex NE, DA, and 5-HT levels were detected. Results Activity of AchE was much higher (P<0.05), but level of ChAT, NE, DA and 5-HT in model group were significantly downregulated compared with normal group (P<0.05). After treated with Kangshuaiyizhi capsule, activity of AchE was downregulated, ChAT, NE, DA and 5-HT levels were significantly upregulated (P <0.01, P <0.05). Conclusion Kangshuaiyizhi capsule can regulate cholinergic and monoamine neurotransmitter levels in the brain tissue of aging model rats, and play a very important role in protecting cerebral function.

ObjectiveTo explore the effects of different doses of PcTx1,a specific blocker of acid-sensing ion channel 1a,on global cerebral ischemia/repedfusion (I/R) injury in rats,MethodsSixty adult male Sprague Dawley rats (weighing 250-300 g) were randomly divided into 6 grups ( n =10 each):sham operation group (group S),I/R group,different doses of PcTx1 ( 10 ng/ml,group P1 ; 25 ng/ml,group P2 ; 50 ng/ml,group P3 ;and 500 ng/ml,group P4 ) groups.Global cerebral ischemia was induced by the modified procedure of Pulsinelli 4-vessel occlusion.In groups P1,P2,P3 and P4,different doses of PcTx1 ( 10,25,50 and 500 ng/ml),6 μl each,were respectively injected into the lateral cerebral ventricle at the initiation of reperfusion,while equal volume of double distilled water was injected instead in group I/R.Six rats in each group were sacrificed at 24 h of reperfusion,and the brains were immediately removed,Thereafter,the contents of malondialdehyde (MDA),reduced glutathione (GSH) and ritric oxide (NO),the activities of constitutive NO synthase (eNOS) snd inducible NO synthase (iNOS) were detected in hippocampus.Four rats in each group were sacrificed at 72 h of reperfusion,and hematoxylin and eosin staining was used to observe the pathomorphological changes of the hippocampal neurons.ResultsCompared with group S,the other groups showed decreases in the contents of GSH,while increases in the contents of MDA and NO and the activities of cNOS and iNOS ( P ＜ 0.05 or 0.01 ).The contents of GSH increased,while the contents of MDA and NO and the activities of cNOS and iNOS decreased in groups P2,P3 and P4 compared with group I/R ( P ＜ 0.05 or 0.01).Compared with group P1,the contents of GSH increased,the contents of MDA and the activities of cNOS decreased in groups P2,P3 and P4,and the contents of NO and the activities of iNOS decreased in groups P3 and P4 ( P ＜ 0.05 or 0.01 ).Compared with group P2,the activities of iNOS decreased in groups P3 and P4(P ＜ 0.05 or 0.01).The damage to neurons in hippocampal CAI was severe in groups I/R and P1,but it was attenuated in groups P3 and P4.ConclusionPcTx1 25,50 and 500 ng/ml (6 μl)injected into lateral cerebral ventricle can attenuate global cerebral I/R injury in rats,and the dose 50 ng/ml (6 μl) is more suitable.

Objective Studies show that the abnormal ex-pressions of APC and survivin play important roles in the development and progression of colon cancer .Survivin shRNA and APC double gene co-expression lentiviral vector was constructed to observe whether it could be successfully expressed in HT-29 colon cancer cells and whether it could impact on colon cancer cell apoptosis . Methods We selected the best shRNA interference fragment from the construc-tion of three pairs of complementary shRNA fragments and connected it with the effective fragment of APC ( aa1020-1698 ) to construct a double gene co-expression lentiviral vector .HT-29 cells were divid-ed into experimental group , empty loading group and blank group .HT-29 cells were observed by fluorescence microscopy after infec-tion.Survivin and APC expression levels were observed by real time PCR and western blot .Apoptosis was detected by caspase-3 activi-ty assay. Results ①We successfully constructed three pairs of shRNA sequences and proved that they had no human gene homolo -gous to the rest.Real time PCR analysis showed that the best sequence was shRNA 3.②After the sequence alignment of constructed shRNA vectors, three pairs of shRNA sequences were completely the same with the first designed sequence .Green fluorescence was observed in HT-29 cells by fluorescence microscope .The survivin content in experiment group (31.71 ±1.49) was significantly de-creased compared with empty loading group (100 ±0) and blank group(95.12 ±2.15)(P<0.05).The expression level of APC mR-NA was significantly increased compared with empty loading group (0 ±0) and blank group(0.51 ±0.15)(P<0.05).③The relative ratio of apoptosis in experiment group (0.573 ±0.050) was significantly increased compared with empty loading group (0.390 ± 0.040) and blank group(0.407 ±0.030)(P<0.05). Conclusion We have successfully constructed survivin-shRNA-APC double gene co-expression lentiviral vector which can be successfully expressed in HT-29 colon cancer cells , providing references for subse-quent gene therapy by the use of the carrier .

Objective Post-transcription RNA interference (RNAi) is more and more widely applied in multigene therapy.This study aimed to establish an subcutaneous xenotransplanted tumor (SXT) model of human HT-29 colon carcinoma in nude mice and investigate the effects of the survivin shRNA-APC double gene co-expression lentiviral vector on the growth of SXT.Methods Thirty-five nude mice were equally divided into five groups, double-gene survivin shRNA, survivin shRNA, APC, empty vector, and blank, and injected into the left anterior axillary with respective stably transfected cell lines and human HT-29 colon carcinoma cells, all at 2×106/mL, to establish an SXT model of human HT-29 colon carcinoma.The inhibition rate of tumor growth was calculated by measuring the size and weight of the SXT, the expressions of survivin mRNA and protein in the tumor tissue detected by real time PCR and immunohistochemistry respectively, and the apoptosis of the HT-29 colon carcinoma cells determined by TUNEL.Results The mean size and weight of the SXT were significantly reduced in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups as compared with the blank and empty vector groups (P<0.05), though increased in the survivin shRNA and APC groups in comparison with the double-gene group (P<0.05).The expressions of survivin mRNA and protein in the tumor tissue were remarkably lower in the double-gene survivin shRNA-APC, survivin shRNA, and APC groups than in the blank and empty vector groups (P<0.05), even lower in the double-gene group than in the survivin shRNA, and APC groups (P<0.05).The apoptosis rate of the HT-29 colon carcinoma cells was markedly up-regulated in the double-gene survivin shRNA-APC ([56.72±3.17]%), survivin shRNA ([33.64±2.03]%), and APC groups ([31.19±1.79]%) as compared with the blank ([9.89±0.31]%) and empty vector groups ([10.06±0.43]%) (P<0.05), even more significantly in the double-gene than in the survivin shRNA and APC groups (P<0.05).Conclusion The survivin shRNA-APC double gene co-expression lentiviral vector can reduce the expression level the survivin gene, promote the apoptosis of colon carcinoma cells, and suppress the growth of the subcutaneous xenotransplanted tumor.

Objective To compare with the effect of high-frequency repetitive transcranial magnetic stimulation (rTMS) between the left and the right prefrontal on refractory negative symptoms and serum brain-derived neurotrophic factor (BDNF). Methods 80 hospitalized schizophrenics with refractory negative symptoms were divided into study group (n=40) and control group (n=40) randomly. Both groups were received 4-week treatment of 10 Hz rTMS. The stimulus lo?cation of the study group was the left prefrontal, and the control group was the right prefrontal. The type and dose of anti?psychotics remained unchanged during the treatment. The evaluation of positive and negative symptom scale (PANSS) and the measurement of BDNF concentration before treatment and after 4 weeks treatment was analyzed. Results Com?pared with before treatment, the total score of PANSS after treatment significantly decreased (P<0.05) both in the study group [(71.2±13.8) vs. (63.3±11.4)] and the control group [(70.3±13.4) vs. (63.7±12.2)]. The score of negative symptoms in the study group decreased [(22.8±6.6) vs. (18.4±5.9), P<0.01]. The BDNF concentration increased in the study group ](6.78±2.16) vs. (8.74±2.76)] and the control group [(6.83±2.32) vs. (8.66±2.70)]. Conclusion 10Hz rTMS on the left pre?frontal combined with drugs are helpful to improve the refractory negative symptom of the patients with schizophrenia. Stimulation on both left and right prefrontal lobe could increase serum BDNF concentration.

Objective To investigate the effect of Yiqi Jianpi herb on content of NPY, VIP and expression of Mapk14 mRNA of rats with spleen-qi deficiency. Methods Rats were randomly divided into normal groups, model group (observed on 7 d, 14 d and 21 d), treatment group of Yiqi Jianpi herb, 10 rats in each group. The spleen-qi deficient model rats were made by rhubarb, exhaustive and hungry method, and treatment group was treated with Sijunzi decoction (20 g/kg) for 21 d, then the content of NPY, VIP in serum and small intestine were evaluated with radioimmunoassay, and expression of Mapk14 mRNA in small intestine tissue was evaluated with real time fluorescent quantitative PCR. Results In model group, content of NPY was much lower than that of normal group (P<0.05), content of VIP and relative expression of Mapk14 mRNA in small intestine tissue were much higher than that of normal group (P<0.05), the difference was obvious in 21 d group. Compared with model 21 d group, content of NPY was increased (P<0.05), content of VIP and relative expression of Mapk14 mRNA in small intestine tissue were decreased (P<0.05) in treatment group. Conclusion Yiqi Jianpi herb has functions of adjusting NPY, VIP secretion and inhibiting abnormal expression of Mapk14 mRNA.

Objective To observe the dynamic changes of ghrelin of spleen-qi-deficiency rats and the intervention effects ofSijunzi Decoction.Methods Experimental rats were randomly divided into normal control group, model group andSijunzi Decoction group. Except for normal control group, spleen-qi-deficiency model was copied through the two-factor methods of breaking qi by bitter cold and swimming exhausted. Meanwhile,Sijunzi Decoction group was given 20 g/(kg?d)Sijunzi Decoction intervention. The activities of GAS, MTL, SS and VIP at different time points (14, 21, 28 d) in intestine and serum were detected by ELISA and RIA. At the same time the intervention effect of Sijunzi Decoction was studied.Results Compared with normal control group, GAS and MTL in intestine and serum of model rats decreased, while SS and VIP increased (P<0.05,P<0.01). Compared with model group, GAS and MTL in intestine and serum of rats in theSijunzi Decoction group increased, while SS and VIP decreased (P<0.05,P<0.01).Conclusion Ghrelin in intestine and serum of spleen-qi-deficiency rats shows dynamic coincidental changes.Sijunzi Decoction can treat spleen qi deficiency by regulating the activities of rat ghrelin.

OBJECTIVETo observe the dynamic time-phase expressions of key genes of brain-gut CaM signal pathway of spleen Qi deficiency rats and the intervention effect of Sijunzi decoction.

METHODMale Wistar rats were randomly divided into the normal control group, model 14 d, 21 d, 28 d groups, and Sijunzi decoction 14 d, 21 d, 28 d groups. Except for the normal control group, the remaining groups were included into the spleen Qi deficiency model with the bitter cold breaking Qi method (ig 7.5 g · kg⁻¹ · d⁻¹ of Rheum officinale, Fructus aurantii immaturus, Magnolia officinalis preparation) and the exhaustive swimming method. On the 7th day after the modeling, the Sijunzi decoction groups were orally administered with Sijunzi decoction 20 g · kg⁻¹ · d⁻¹. The expressions of key genes CaM/CaMK II of CaM signaling pathway in hippocampus and intestine at different time points by immunohistochemical method and Western blot. At the same time, the intervention effect of Sijunzi decoction on spleen Qi deficiency rats and its mechanism were analyzed.

RESULTSpleen Qi deficiency rats showed higher intestinal CaM/CaMK II expression and lower hippocampus CaM/CaMK II expression than normal rats (P < 0.05, P < 0.01). After the treatment of Sijunzi decoction, spleen Qi deficiency rats showed reduction in intestinal CaM/CaMK II expression and increase in hippocampus CaM/CaMK II expression (P < 0.05, P < 0.01).

CONCLUSIONThe formation of spleen Qi deficiency syndrome may be related to the high expression of CaM/CaMK II in small intestine tissues and its low expression in hippocampus tissues. Sijunzi decoction may achieve the therapeutic effect in spleen Qi deficiency syndrome by reducing the CaM/CaMK II expression in intestinal tissues and increasing it in hippocampus tissues.

Animals ; Brain ; drug effects ; enzymology ; metabolism ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Calmodulin ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Intestines ; drug effects ; enzymology ; metabolism ; Male ; Qi ; Rats ; Rats, Wistar ; Spleen ; drug effects ; Splenic Diseases ; drug therapy ; enzymology ; genetics ; metabolism

Objective This study aimed to explore issues about dysandia female during pregnancy,peripartum and contraception and to supply reference for their reproductive health improvement. Methods Purposive sampling strategy was used. With the aid of an interview guide, 12 dysaudia women who met inclu-sion and exclusion criteria were invited to be interviewed. Interviews were recorded, validated and then ana-lyzed. Results Four themes were generated, limited knowledge, passive information acquisition, total depen-dence on family and reluctant to talk about contraception. Total dependence on family was noticeable during pregnancy and peripartum. Though little was known about health protection related to pregnancy, peripartum and contraception, participants were passive at seeking for relevant information. Conclusions Deaf women may experience more threats to reproductive health in relation to pregnancy, peripartum and contraception that they require more guidance and assistance.

OBJECTIVETo investigate the neuro-protective effects of baicalin in Wistar rats with focal cerebral ischemic reperfusion injury.

METHODSNinety adult male Wistar rats weighing 320-350 g were randomly divided into the following groups (n=5): (a) sham control group; (b) vehicle group, subjected to middle cerebral artery occlusion and received vehicle intraperitoneally; (c-e) baicalin groups, which were subjected to the middle cerebral artery occlusion and treated with baicalin 25, 50 and 100 mg/kg, respectively. The neurological scores were determined at postoperative 1, 3 and 7 d after the treatment. The expression of protease-activated receptor-1 (PAR-1), PAR-1 mRNA and Caspase-3 were determined using Western blot, reverse transcription polymerase chain reaction (RTPCR) analysis and immunohistochemistry, respectively.

RESULTSSignificant decrease was noted in the neurological score in the baicalin group compared with that of the vehicle group (P<0.01). Additionally, down-regulation of PAR-1 mRNA, PAR-1 and Caspase-3 was observed in the baicalin groups compared with those obtained from the vehicle group (P<0.01). Compared with the low-dose baicalin group (25 mg/kg), remarkable decrease was noted in neurological score, and the expression of PAR-1 mRNA, PAR-1 as well as Caspase-3 in the high-dose group (P<0.05).

CONCLUSIONBaicalin showed neuro-protective effects in focal cerebral ischemic reperfusion injury through inhibiting the expression of PAR-1 and apoptosis.

Animals ; Apoptosis ; drug effects ; Brain Ischemia ; complications ; drug therapy ; genetics ; pathology ; Caspase 3 ; metabolism ; Flavonoids ; administration & dosage ; pharmacology ; therapeutic use ; Gene Expression Regulation ; drug effects ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Receptor, PAR-1 ; antagonists & inhibitors ; genetics ; metabolism ; Reperfusion Injury ; complications ; drug therapy ; genetics ; pathology