This study is to investigate the effects of aqueous extract of Schisandra chinensis Baill (WWZ), kadsurin, schisandrin A, schisandrin B and schisandrol B on rat hepatic CYP3A. Rats received a daily gavage of aqueous extract of WWZ for different times. The livers were harvested after gavage and subjected to microsome preparation. Microsomal CYP3A activity was determined by measuring the amount of the metabolite of testosterone (6 beta-hydroxytestosterone) with HPLC. Aqueous extract of WWZ, kadsurin and schisandrin A were incubated with microsomes obtained from rat. Microsomal CYP3A activity was determined by HPLC. Primary hepatocytes were separated and extracted from rat, then were treated with aqueous extract of WWZ, schisandrin A, schisandrin B and schisandrol B. Then, the expression of CYP3A1 mRNA was analyzed by RT-PCR. As for the in vivo assay, aqueous extract of WWZ significantly inhibited the enzyme activity of CYP3A after 12 h gavage. The inhibitory effect was converted to inductive effect after 3-day gavage. Aqueous extract of WWZ could induce the enzyme activity of CYP3A after 6-day gavage. Aqueous extract of WWZ and kadsurin showed a dose-dependent inhibition of CYP3A (IC50 of 487.8 microg mL(-1) and 6.2 micromol L(-1), separately). In rat primary hepatocytes, aqueous extract of WWZ (2.5 mg mL(-1)), schisandrin A (0.1 micromol L(-1)), schisandrin B (0.1 micromol L(-1)) and schisandrol B (10 micromol L(-1)) increased significantly the expression of CYP3A1 mRNA by 23%, 55%, 42% and 27%, respectively. Aqueous extract of WWZ could show dual effect on the enzyme activity of CYP3A in rat in vivo. Meanwhile, kadsurin showed a dose-dependent inhibition of the enzyme activity of hepatic CYP3A in vitro. And schisandrin A, schisandrin B and schisandrol B showed significant inductive effect on the expression of rat CYP3A1 mRNA.

Background and purpose:One of the reasons why cancer cells are resistant to chemotherapy is the existence of cancer stem cells. The purpose of this study was to investigate the chemoresistance of CD44+/CD24+ Siha cells to cisplatin and its mechanisms.Methods:Siha cells were cultivatedin vitro. The CD44+/CD24+ Siha cells were sorted out by fluorescence activated cell sorter (FACS) andin vitro proliferation was detected by MTT assay after treatment with the different concentrations of cisplatin. The cell apoptosis rate was detected by flow cytometry after 10 μg/mL cisplatin acted on CD44+/CD24+ Siha cells for 24, 48 and 72 h. The relative mRNA and protein expressions of Bcl-2, Oct-4 and ABCG2 were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Results:The survival rates of CD44+/CD24+ Siha cells treated with different concentrations of cisplatin (0.1, 1, 5, 10, 15 and 20 μg/mL) were higher than those of their parental Siha cells [(88.42±1.51)%vs (92.87±1.5)%, (79.94±1.05)%vs (84.72±1.09)%, (69.78±0.81)%vs (75.13±2.86)%, (58.97±0.70)%vs (65.79±2.71)%, (49.60±0.88)%vs (52.10±0.52)%, (45.13±0.69)%vs (48.84±1.02)%,P<0.05]. Compared with their parental Siha cells, the apoptosis rates of CD44+/CD24+ Siha cells were lower after 10 μg/mL of cisplatin acting on them for 24, 48 and 72 h, respectively [(3.05±0.16)%vs (5.17±0.27)%, (17.94±2.02)%vs (32.60±4.28)% and (40.14±3.01)%vs (56.62±5.32)%,P<0.05]. The results from both qRT-PCR and Western blot indicated that Oct-4, ABCG2 and Bcl-2 were highly expressed on CD44+/CD24+ Siha cells. A significant difference was found in Oct-4, ABCG2 and Bcl-2 expression between CD44+/CD24+Siha cells and their parental cells (P=0.015<0.05).Conclusion:CD44+/CD24+ Siha cells could be resistant to apoptosis induced by cisplatin and expressed high levels of cancer stem cell markers such as Oct-4 and ABCG2. This study lays the basis for useful isolation and further targeted therapy of cervical cancer stem cells.

Objective To explore whether CD44 +/CD24 + expressing cervical cancer cells are resistant to X-ray irradiation and investigate the underlying mechanism.Methods Cervical cancer cell line (Siha) was cultured in vitro and the CD44 +/CD24 + expressing cells were sorted with a flow cytometer.The cells were irradiated with 8,16 and 30 Gy of 6 MV X-rays.Colony formation test was used to evaluate the radiosensitivity of CD44 +/CD24 + expressing cervical cancer cells.Cell morphology was observed by electronmicroscopy,cell apoptosis was analyzed with a flow cytometer and also verified with a DNA ladder assay.Gene expression was determined by RT-PCR.Results After radiation,the ratio of CD44 +/CD24 + cells significantly increased.Compared to Siha cells,the radiosensitivity of CD44 +/ CD24 + cells decreased (t =93.99-400.45,P ＜0.05),and the expressions of bcl-2,survivin and Oct4 mRNA increased in CD44 +/CD24 + cells (t =221.35,941.65,82.27,P ＜0.01).Both apoptotic body and specific DNA ladder pattern were observed in cells but not in the CD44 +/CD24 + Sihacells which had no obvious morphological changes of apoptosis.Conclusions The CD44 +/CD24 + expressing cervical cancer cells are resistant to X-rays due to expression of anti-apoptosis factors.

Objective This study was designed to observe the impact of cigarette cessation on anxiety and depression in patients with coronary heart disease (CHD).Methods A total of 690 cigarette smoking patients with CHD identified by coronary angiography (CAG) were included and analyzed in the study.The mental state were scored with Hamilton anxiety (HAMA) and depression (HAMD) scale both on admission and at 6-month follow-up.The patients were divided into two groups based on the cigarette cessation.The score of mental state between the two groups were compared.The patients were treated with percutaneous coronary intervention (PCI),coronary artery bypass grafting (CABG),or medicine therapy (MT).Results The clinic data and score of mental state were similar at the time of admission (HAMA:10.66±5.53 vs 11.09 ±5.61,P =0.311;HAMD:29.81 ± 10.13 vs 28.94 ± 10.22,P =0.266 4) between the two groups.After 6 months,the proportions of subjects in smoking cession group with anxiety (24.2％ vs 32.3％,P ＜0.05),depression (18.0％ vs 27.5％,P ＜0.05),and anxiety and depression (7.0％ vs 16.2％,P ＜0.001) decreased significantly compared with those in smoking group irrespective of the treatment strategy.Both the HAMA and HAMD scores were lower in smoking cessation group (HAMA:9.83±3.40;HAMD:24.91 ±7.90) than in smoking group (HAMA:10.98 ±4.87;HAMD:27.70 ± 11.16) (all P ＜ 0.001).Conclusions Smoking cessation is good for the relief of anxiety and depression in CHD patient.

AIM: To investigate the therapeutic effects of Zhuyejiao tablets (Tab.ZYJ) on chronic pelvic inflammation (CPI) induced by coliform in rats. METHODS: The CPI model was made by injecting coliform O-B_4 standard strains in the uterus of rat. Animals were randomly divided into six groups and drugs were administered for 21 days, bid, respectively. The immune function of animals was measured and the uterus was pathologically observed. RESULTS: The level of serum agglutinin and lymphocyte transformation index markedly increased in all ZYJ groups. Morphological investigation also revealed the alleviation of inflammation in ZYJ groups. CONCLUTION: ZYJ has therapeutic effects on chronic pelvic inflammation in rats.

ObjectiveTo evaluate the effects of combined vagus nerve electric stimulation postconditioning and limb remote ischemic postconditioning on myocardial ischemia-reperfusion (I/R) injury in rats.Methods One hundred male SD rats aged 8 weeks weighing 250-350 g were randomly allocated into 5 groups ( n =20 each):sham operation group (group S); I/R group; vagus nerve electric stimulation postconditioning group (group POES) ; limb remote ischemic postconditioning group (group RP) and vagus nerve electric stimulation postconditioning + limb remote ischemic postconditioning group (group POES-RP).Myocardial I/R was induced by occlusion of left anterior descending branch (LAD) of coronary artery for 30 min followed by 120 min reperfusion in groups I/R,POES,RP and POES-RP.In groups POES and POES-RP,right cervical vagus nerve trank was stimulated for 30 min with continuous electric rectangular pulses (2 ms,10 Hz) starting from 15 min of myocardial ischemia.The voltage of the pulses was adjusted to decrease HR by 10％ of the baseline HR before stimulation.In groups RP and POES-RP the animals underwent 10 min ischemia of bilateral hind limbs starting from 20 min of myocardial ischemia.Arterial blood samples were collected from 10 rats in each group at 120 min of reperfusion for determination of serum concentrations of cTnI,CK-MB,TNF-α,high mobility group box 1 protein (HMGB1),intercellular adhesion molecule-1(ICAM-1),IL-1,IL-6 and IL-10 (by ELISA).The animals were then sacrificed and myocardial infarct size was measured by Evans blue and TTC staining.Another 10 rats were sacrificed at 120 min of reperfusion for determination of myocardial contents of TNF-α,HMGB-1,ICAM-1,IL-1,ID6 and IL-10 (by ELISA).ResultsI/R induced myocardial infarct and significantly increased serum concentrations of cTnI,CK-MB,TNF-α,HMGBi,ICAM-1,IL-1 and IL-6 and increased myocardial contents ofTNF-α,HMGB1,ICAM-1,IL-1,IL-6 and IL-10 in both ischemic and non-ischemic regions in group I/R as compared with group S.Vagus nerve electric stimulation postconditioning,limb remote ischemic postconditioning and vagus nerve electric stimulation postconditioning + limb remote ischemic postconditioning significantly decreased myocardial infarct size and serum concentrations of cTnI,CK-MB,TNF-α,HMGB1,ICAM-1,IL-1 and IL-6 and decreased myocardial contents of TNF-α,HMGB1,ICAM-1,IL-1,IL-6 in groups POES,RP and POES-RP as compared with group I/R.Compared with group I/R,myocardial IL-10 content in both ischemic and non-ischemic regions was significantly increased in groups POES and POES-RP.Compmared with group POES,myocardial infarct size,serum concentrations of cTnI,CK-MB,TNF-α,ICAM-1 and myocardial contents of ICAM-1 and IL-6 in ischemic region were significantly decreased,while myocardial content of IL-10 in non-ischemic region was increased in group POES-RP.Compared with group RP,myocardial infarct size,serum concenuations of cTnI,CK-MB,TNF-α,HMGB1,ICAM-1,IL-1,IL-6 and myocardial contents of TNF-α,ICAM-1,IL-1 and IL-6 in ischemic region were significantly decreased,myocardial content of IL-10 in ischemic region was increased and HMGB1,ICAM-1,IL-1 and IL-6 contents were decreased,IL-10 content was increased in myocardial of ischemic region in group POES-RP.ConclusionMyocardial I/R injury is attenuated and myocardial protection is enhanced by combination of vagus nerve electric stimulation postconditioning and limb remote ischemic postconditioning in rats by inhibiting inflammatory response in rats.

Objective To investigate the role of reactive oxygen species (ROS) in the reduction of myocardial ischemia-reperfusion (I/R) injury by fentanyl postconditioning and remote limb ischemic postconditioning in rats.Methods Sixty-three male Sprague-Dawley rats,aged 8 weeks,weighing 250-350 g,were equally and randomly allocated into 7 groups:sham operation group (group S),group I/R,fentanyl postconditioning group (group F),remote limb ischemic postconditioning group (group R),ROS scavenger N-(2-Mercaptopropionyl) glycine (MPG) group (group M),MPG + fentanyl postconditioning group (group MF),and MPG + remote limb ischemic postconditioning group (group MR).Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 180 min of reperfusion.In group S the anterior descending branch was only exposed but not ligated.MPG 5 mg/kg was infused intravenously from 5 min before ischemia to 15 min of reperfusion in groups M,MF and MR,while the equal volume of normal saline was given in the other four groups.In groups F and MF,fentanyl 30 μg/kg was injected intravenously at 15 min of myocardial ischemia.In groups R and MR,the animals underwent 10 min ischemia of bilateral hind limbs starting from 15 min of myocardial ischemia.Arterial blood samples were taken at 180 min of reperfusion to determine the serum cardiac troponin I (cTnI) concentration.The rats were then sacrificed.The infarct size was measured by TTC.Results Compared with group S,the serum cTnI concentration and infarct size were significantly increased in the other six groups (P ＜0.05).Compared with group I/R,no significant change was found in the serum cTnI concentration and infarct size in M group,and the serum cTnI concentration and infarct size were significantly decreased in F and R groups (P ＜ 0.05).There was no significant difference in the serum cTnI concentration and infarct size between MF group and F group (P ＞ 0.05).The serum cTnI concentration was significantly higher and the infarct size was larger in group MR than in group R (P ＜ 0.05).Conclusion ROS is involved in the reduction of myocardial I/R injury by remote limb ischemic postconditioning in rats,but not in the myocardial protection provided by fentanyl postconditioning.

Objective To evaluate the roles of PI3K/Akt and JAK/STAT signal transduction pathways in reduction of myocardial ischemia/reperfusion (I/R) injury by postconditioning with α subunit-containing nicotinic acetylcholine receptor (α7nAChR) agonist in rats.Methods Sixty Sprague-Dawley rats,weighing 290-320 g,were randomly divided into 4 groups (n =15 each):I/R group,ischemic preconditioning group (IPC group),ischemic postconditioning group (IPOC group) and postconditioning with specific α7nAChR agonist PNU282987 group ( PNU group ).Myocardial I/R was produced by 30 min occlusion of left anterior descending coronary artery followed by 180 min reperfusion in the 4 groups.The animals were subjected to 3 cycles of 5 min myocardial ischemia and 5 min reperfusion before 30 min myocardial ischemia in IPC group.The animals underwent 3 cycles of 10 s myocardial ischemia at 5 s intervals before 180 min reperfusion in group IPOC.PNU282987 2.4 mg/kg was injected intraperitoneally immediately before the reperfusion.At 60 min of reperfusion,5 rats in each group were sacrificed and the hearts were removed to determine the expression of Akt and STAT3 mRNA,phosphorylated Akt (p-Akt) and phosphorylated STAT3 (p-STAT3) in myocardial tissues.The left 10 rats in each group were sacrificed at 180 min of reperfusion and the hearts were removed to measure the infarct size.Results Compared with I/R group,the expression of STAT3 mRNA and p-Akt was significantly up-regulated in IPC group,and the expression of p-Akt and p-STAT3 was significantly up-regulated in IPOC group ( P ＜ 0.05).The infarct size was significantly reduced in IPC,IPOC and PNU groups compared with I/R group ( P ＜ 0.05 ).Conclusion The mechanism by which α7nAChR agonist postconditioning reduces myocardial I/R injury is not related to PI3K/Akt and JAK/STAT signal transduction pathways in rats.

Objective To explore the protective effect and mechanism of glycyrrhizic acid on hippocampal neuron injury in epileptic rat models. Methods The epileptic rat models were established by lithium and pilocarpine kindling. The successful models were randomly divided into epilepsy group and glycyrrhizic acid group. In addition, the rats without any treatment were used as the normal control group, with 12 rats in each group. Hippocampal neuron injury and apoptosis were detected by Nissl and TUNEL staining. Mitochondrial membrane potential in the hippocampal neurons of rats was detected by JC-1 method. The activity of aspartic acid protein hydrolase 3 (caspase 3) and caspase 9 was detected by colorimetric assay. The expressions of cleaved-caspase 3, 9, B lymphocyte tumor-2 (Bcl-2), Bcl-2 related X protein (Bax), cytochrome C (CytC) and apoptotic protease-activating factor (Apaf-1) protein in the hippocampal tissues of rats were detected by western blot assay. Results Compared with the normal group, the number of neurons was reduced (P＜0. 01), the number of TUNEL-positive cells was increased (P＜ 0. 01), mitochondrial membrane potential was decreased (P＜ 0. 01), caspase 3 and 9 activity was increased (P＜ 0. 01), the expressions of Bcl-2 and mitochondrial CytC were down-regulated (P＜ 0. 01), the expressions of Bax, cytoplasm CytC and Apaf-1 were up-regulated (P＜ 0. 01) in the epilepsy group. Compared with the epilepsy group, the number of neurons was increased (P ＜ 0. 01), the number of TUNEL-positive cells was reduced (P＜ 0. 01), mitochondrial membrane potential was increased (P＜ 0. 01), caspase 3 and 9 activity was decreased (P ＜ 0. 01 ), the expressions of Bcl-2 and mitochondrial CytC were up-regulated (P ＜0. 01), the expressions of Bax, cytoplasm CytC and Apaf-1 were down-regulated (P ＜ 0. 01) in the glycyrrhizic acid group. Conclusions These results suggest that glycyrrhizic acid can inhibit the hippocampal neuron injury in epileptic rats by blocking the mitochondrial pathway.

Objective To explore the effect of low-level laser therapy (LLLT)applied to the quadriceps muscle on the recovery of exhausting-cycling-exercise-induced fatigue.Methods According to a randomised,double-blind and crossover design,16 healthy male students were randomly assigned to an LLLT-1,LLLT-3,LLLT-5 and a placebo group,and received LLLT for 300 s at the dosage of 0.06 J· cm-2,0.18 J·cm-2,0.3 J·cm-2 and 0 to the bilateral rectus femoris after the exhausting-cycling-exercise-induced fatigue.The blood lactate(BL),heart rate(HR),rated perceived exertion(RPE)and visual analogue scale(VAS)were assessed before the exercise,immediately after exercise,10 and 20 min after exercise,as well as immediately after the first Wingate(WG)test,5 and 30 min after the WG test.Meanwhile,the second WG test was performed 40 min after the first WG test.Results The average HR value of LLLT-1 group was significantly lower than the placebo group at 10 min after exercise(P＜ 0.05)and immediately after the WG test(P＜0.01),while that of LLLT-3 and LLLT-5 groups was significantly lower than the placebo group immediately and 5 min after the WG test(P＜0.01).Compared to the placebo group,the average BL of LLLT-1,LLLT-3 and LLLT-5 groups was significantly lower 10 min after exercise(P＜0.05 for all)and that of LLLT-5 group was also significantly lower 30 min after the first WG test(P＜0.05).However,the average blood glucose of LLLT-5 group was significantly higher than the placebo group right after the first WG test(P＜0.05).Moreover,significant increase was observed in the mean(P=0.002)and peak power(P=0.006)at the second WG test and the mean(P=0.048) power at the first WG test of LLLT-3 group,compared to the placebo group.Conclusion LLLT applied to quadriceps muscles after exhausting exercise may enhance recovery,and LLLT at the dose of 0.18 J·cm-2 is of the best effect.