Objective To study the cardiomyocyte and renal cell apoptosis induced by overtraining, and the preventive effects of anisodamine against the adverse effects. Methods The rats were forced to swim till exhaustion to reproduce the animal model of overtraining. The animals were randomly divided into control group, exhausted group and anisodamine group. The exhausted group, depending on the recovery time after exhaustion, was divided again into exhaustion subgroup, 6h after exhaustion subgroup, and 24h after exhaustion subgroup. The animals in the anisodamine group received intraperitoneally 10mg/kg of anisodamine before the swimming overtraining, and divided again into 6h after anisodamine injection subgroup and 24h anisodamine injection subgroup. The cardiomyocyte and renal cell apoptosis was observed by the method of TUNEL, image analysis and flow cytometry. Results It was revealed by TUNEL that the number of apoptotic cardiomyocytes was increased after exhaustive swimming, especially in the 6h after exhaustion subgroup. The number of apoptotic cells was also increased in kidney of 0h, 6h and 24h after exhaustion subgroups, especially in 24h after exhaustion subgroup. The apoptosis ratio was also increased significantly in 0h, 6h and 24h after exhaustion subgroups as demonstrated by flow cytometry. Compared with the rats of exhausted group, the number of apoptotic cells in heart and renal tissue was decreased remarkably after anisodamine injection. Conclusion The apoptosis of cardiomyocyte and renal cell could be induced by exhaustive swimming. Compared with kidney, heart injury recovered more quickly. Anisodamine had the preventive effect on the injury to heart and kidney in exhausted rats.

Objective To evaluate the role of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2) in 17β estradiol-induced inhibition of propofol-caused neuroapoptosis in the hippocampus of newborn rats.Methods Seventy-eight male Sprague-Dawley rats,aged 7 days,weighing ll-18 g,were divided into 6 groups (n =13 each) using a random number table:dimethyl sulfoxide (DMSO) group,fat emulsion group (group F),17β estradiol group (group E),propofol group (group P),propofol plus 17β estradiol group (group PE) and propofol plusl7β estradiol plus mitogen-activated protein kinase kinase 1/2 inhibitor U0126 group (group PEU).17β estradiol 600 μg/kg was injected subcutaneously every 24 h for 7 consecutive days in group E,and the equal volume of DMSO was given instead in group DMSO.Propofol 75 mg/kg was injected intraperitoneally every 24 h for 7 consecutive days in group P,and the equal volume of fat emulsion was injected instead in group F.Propofol 75 mg/kg was injected intraperitoneally,and 17β estradiol 600 μg/kg was injected subcutaneously every 24 h for 7 consecutive days in group PE.Propofol 75 mg/kg was injected intraperitoneally,17β estradiol 600 μg/kg was injected subcutaneously,and U0126 10 mg/kg was injected intraperitoneally every 24 h for 7 consecutive days in group PEU.At 15 min after the last injection,3 rats in each group were randomly selected,and arterial blood samples from the cardiac apex were collected for determination of arterial oxygen partial pressure.The animals were sacrificed at 24 h after the last injection for determination of the expression of activated caspase-3 (by immunohistochemistry) and p-ERK1/2 (by Western blot).Results There was no significant difference in arterial oxygen partial pressure between the six groups (P＞0.05).Compared with group F,the expression of activated caspase-3 was significantly up-regulated,and the expression of p-ERK1/2 was downregulated in group P (P＜0.05).Compared with group P,the expression of activated caspase-3 was significantly down-regulated,and the expression of p-ERK1/2 was up-regulated in group PE (P＜0.05).Compared with group PE,the expression of activated caspase-3 was significantly up-regulated,and the expression of p-ERK1/2 was down-regulated in group PEU (P＜0.05).Conclusion The mechanism by which 17β estradiol inhibits propofol-caused neuroapoptosis in the hippocampus is related to up-regulation of the expression of p-ERK1/2 in newborn rats.

Objective To investigate the effect of penehyclidine hydrochloride ( PHC) pretreat?ment on nuclear factor erythroid 2?related factor 2∕heme oxygenase?1 ( Nrf2∕HO?1) signaling pathway in re?nal tissues of rats with rhabdomyolysis?induced acute kidney injury ( AKI) . Methods Thirty?six pathogen?free male Sprague?Dawley rats, weighing 200-220 g, were assigned into 3 groups ( n=12 each) using a random number table: control group (group C), group AKI and PHC pretreatment group (group PHC). Rhabdomyolysis was induced by intramuscular injection of 50% glycerol 10 ml∕kg in bilateral hindlimbs. PHC 0?2 mg∕kg was injected intraperitoneally at 30 min before glycerol was injected intramuscularly in group PHC. At 1 and 6 h after glycerol injection, serum was collected for determination of blood urea nitro?gen ( BUN) and creatinine ( Cr) concentrations, and bilateral kidneys were harvested for pathological ex?amination and for determination of HO?1 activity and expression of Nrf2 mRNA and HO?1 mRNA ( by quan?titative real?time polymerase chain reaction) , Nrf2 in nucleoprotein and total protein and HO?1 in total pro?tein in renal tissues ( by Western blot) . The damage to the renal tubules was scored. Results Compared with group C, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly increased, the expression of Nrf2 in nucleoprotein and total protein and HO?1 in total protein was signifi?cantly up?regulated, and HO?1 activity was significantly increased in AKI and PHC groups, the expression of HO?1 mRNA was significantly up?regulated in group AKI, and the expression of Nrf2 mRNA and HO?1 mRNA was significantly up?regulated in group PHC (P<0?01 or 0?05). Compared with group AKI, the BUN and Cr concentrations in serum and renal tubular damage scores were significantly decreased, the ex?pression of Nrf2 in nucleoprotein and total protein and HO?1 in total protein was significantly up?regulated, and HO?1 activity was significantly increased in group PHC ( P<0?01 or 0?05) . Conclusion The mecha?nism by which PHC pretreatment attenuates rhabdomyolysis?induced AKI may be related to activation of Nrf2∕HO?1 signaling pathway in renal tissues of rats.

Objective To evaluate the role of P2X3 receptors in dorsal root ganglion in development of incisional pain in rats.Methods Twenty-four healthy male SD rats weighing 200-220 g were randomly divided into 3 groups(n = 8 each): control group(group C),incisional pain(group IP)and P2X3 receptor antagonist + IP group(group A).In group IP and A,a 1 cm longitudinal incision was made in the plantar surface of left hindpaw according to the method described by Brennan et al.in isoflurane-anesthetized rats.P2X3 receptor antagonist TNP-ATP 200 nmol was injected into the plantar surface of left hindpaw 30 min after plantar incision was made in group A,while equal volume of normal saline was given instead of TNP-ATP in group C and IP.The behavior of the hindpaw of the rats were assessed using cumulative pain score within 1 h after injection.The animals were sacri ficed 2 h after injection and the dorsal root ganglion was removed for determination of P2X3 receptor expression and intracellular Ca2+ concentrations.ResultsThe cumulative pain scores,P2X3 receptor expression and Ca2 + concentrations were significantly higher in group IP and A than in group C(P ＜ 0.05).The cumulative pain scores,P2X3 receptor expression and Ca2+ concentrations were significantly lower in group A than in group IP(P ＜0.05).Conclusion P2X3 receptors in dorsal root ganglion is involved in the development of incisional pain through increasing intracellular Ca2+ concentrations in rats.

Objective To explore the clinical features of diabetic macular edema (DME) with optical coherence tomography (OCT) and correlation with visual function. Methods Forty-nine eyes from 40 patients with DME (DME group) and 31 eyes from 31 patients without DME (control group) were examined with OCT,pattern reversal visual evoked potentials (P-VEP),macular perimetry. According to proliferative diabetic retinopathy (PDR), 49 eyes with DME were divided into group A (without PDR, 30eyes) and group B (with PDR, 19 eyes). Results The retinal macular thickness of central fovea in DME group [(299.25±63.87)μm] was more than that in contol group [(204.35 ± 37.94)μm], visual acuity and macular visual field in DME group were significantly different than those in control group, respectively (P ＜ 0.05). The retinal macular thickness of central fovea,visual acuity and visual field were no significant differences between group A and group B (P＞0.05). OCT macular thickness and visual correlation coefficient was -0.437(P＜ 0.05 ); OCT macular thickness and mean defect correlation coefficient was 0.441(P ＜ 0.05). Conclusions OCT can provide a useful tool for monitoring the occurrence and development of DME, can assess the response to treatment. With increasing of the macular retinal thickness, the visual acuity and macular visual field of visual function are more damaged.

Objective To evaluate the effects of anisodamine on apoptosis in cardiomyocytes and inflammatory response in overtrained rats. Methods Twenty-four male Wistar rats, weighing 200-220 g, were randomly divided into 3 groups ( n = 8 each) : control group (group C) , overtraining group (group O) , anisodamine group (group A) . The model of overtraining-induced acute heart injury was established by exhausting swimming. Anisodamine 10 mg/kg was given intraperitoneally 20 min before overtraining in group A. Blood samples were taken at 6 h after overtraining for measurement of serum CK-MB activity. The rats were then sacrificed and myocardial tissues taken for determination of TNF-α content and NF-κB activity (by immunohistochemistry) . The apoptosis rate was detected by flow cytometry. Results The CK-MB activity, apoptosis rate, TNF-α content and NF-κB activity were significantly higher at 6 h after overtraining in groups O and A than in group C, while lower at 6 h after overtraining in group A than in group O ( P < 0.05) . Conclusion Anisodamine can inhibit apoptosis in cardiomyocytes by reducing inflammatory response in overtrained rats.

Objective To investigate the effects of Inula Britannica on myocardial caspase-3 and cytochrome c ( cyt c) following overtraining-induced acute myocardial injury in rats.Methods Forty-eight male Wistar rats weighing 200-220 g were randomly divided into 3 groups:group control (group C,n =8) ; group exhausting swim (group E,n =24) and group Inula Britannica (group IB,n =16).The animal model of overtraining-indnced acute myocardial injury was developed by exhausting swim.The animals were forced to swim until they were exhausted.The animals sank to the bottom and no righting reflex or escape response was elicited when they were taken out of water in groups E and IB.In group IB oral Inula Britannica 25 ml/kg was given 24 h and immediately before overtraining.Blood samples were taken from inferior vena cava immediately and at 6,24 h after overtraining in group E and at 6,24 h after overtraining in group IB for determination of serum cardiac troponin I (cTnI) concentration (by ELISA).The animals were sacrificed after blood sampling and myocardial specimens were obtained for microscopic examination and determination of caspase-3 and cyt c expression (by immuno-histochemistry).Results Overtraining significantly increased serum cTnI concentration and up-regulated myocardial caspase-3 and cyt c expression in group E as compared with group C.Oral Inula Britannica significantly attenuated overtraining-induced increase in serum cTnI concentration and myocardial caspase-3 and cyt c expression in group IB as compared with group E.Conclusion Inula Britannica can reduce overtraining-induced acute myocardial injury by down-regulating caspase-3 and cyt c expression.

Objective To evaluate the effect of ixeris sonchifolia on mitochondrial transcription factor A (mtTFA) expression following myocardial injury induced by exhausting exercise in rats.Methods Forty male Wistar rats,weighing 200-220 g,were randomly divided into 3 groups using a random number table:control group (group C,n =8),exhausting exercise group (group ES,n =16) and ixeris sonchifolia group (group IS,n =16).The model of myocardial injury was established by exhausting swimming.In IS group,the rats were subjected to exhausting swimming after intraperitoneal ixeris sonchifolia 20 g/kg.In ES and IS groups,8 rats were chosen at 6 and 24 h after exhaustion,and blood samples were taken from the inferior vena cava for determination of serum cardiac troponin I (cTnI) concentrations by ELISA.The animals were sacrificed after blood sampling and myocardial specimens were obtained for microscopic examination and for determination of mtTFA expression by Western blot.Results Compared with group C,the concentration of serum cTnI was significantly increased,and the expression of mtTFA in myocardial tissues was down-regulated at 6 and 24 h after exhaustion in ES and IS groups.Compared with group ES,the concentration of serum cTnI was significantly decreased,and the expression of mtTFA in myocardial tissues was up-regulated at 6 and 24 h after exhaustion in group IS.Conclusion Ixeris sonchifolia can reduce exhausting exercise-induced myocardial injury through up-regulating myocardial mtTFA expression in rats.

Objective To evaluate the effect of anisodamine on endoplasmic reticulum stress during acute kidney injury (AKI) in rats.Methods Forty-two nale Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 3 groups:control group (group C,n =6),AKI group (n =18) and anisodamine group (group AD,n =18).AKI was induced by intramuscular injection of 50％ (v/v) glycerol 10 ml/kg into bilateral hind limbs in groups AKI and AD.In addition,anisodamine 10 mg/kg was injected intraperitoneally 20 min before intramuscular injection of glycerol in group AD.In group C the rats received intramuscular injection of normal saline 10 ml/kg into bilateral hind limbs.Six rats were chosen immediately after injection of normal saline in group C or at 1,6 and 24 h after glycerol injection in groups AKI and AD and then anethetized with intraperitoneal pentobarbital.The animals were sacrificed and kidney specimens were obtained and cut into sections which were stained with hematoxylin and eosin for pathological examination.The pathological changes of the renal tubules were scored.The expression of glucose-regulated protein 78 (GRP78) and oxygen-regulated protein 150 (ORP150) in renal tissues was determined by immuno-histochemistry and Western blot.Results Compared with group C,the pathological scores were significantly increased and the expression of GRP78 and ORP150 was up-regulated at all time points in groups AKI and AD (P ＜ 0.01).Compared with group AKI,the pathological scores were significantly decreased and the expression of GRP78 and ORP150 was down-regulated at all time points in group AD (P ＜ 0.05 or 0.01).Conclusion Anisodamine can ameliorate AKI through inhibiting endoplasmic reticulum stress in renal tubular epithelial cells and decreasing endoplasmic reticulum stress-induced cell apoptosis in rats.

Objective To evaluate the effects of curcumin pretreatment on the expression of Nrf2 protein during ventilator-induced lung injury in rabbits.Methods Twenty-four healthy male New Zealand white rabbits,aged 3-6 months,weighing 2.5-3.0 kg,were randomized into 3 groups (n =8 each) using a random number table:two-lung ventilation (TLV) group; one-lung ventilation (OLV) group; and curcumin pretreatment group (group Cur).In group Cur,curcumin 40 mg/kg (dissolved in 2 ml of 1％ sodium carboxymethylcellulose) was given via a gastric tube into the stomach twice a day for 7 consecutive days starting from 7 days before ventilation,while the equal volume of sodium carboxymethylcellulose was given via a gastric tube instead of curcumin in TLV and OLV groups.All the rabbits were tracheostomized,and a tracheal tube was inserted to perform TLV in TLV group,and a tracheal tube was inserted into the right bronchus to establish OLV in OLV and Cur groups.Volumecontrolled ventilation was used in the three groups and the ventilatory parameters were regulated to maintain SpO2 ＞ 90 ％.Immediately before beginning of ventilation (T0) and at 1,2 and 3 h of ventilation (T1-3),arterial blood samples were obtained for blood gas analysis and determination of PaO2.The oxygenation index was calculated.At the end of ventilation,the rabbits were sacrificed and right lungs were removed for determination of wet/dry lung weight ratio (W/D ratio).The right lower lobe was isolated and puhmonary specimens were obtained for determination of malondialdehyde (MDA) content and superoxide dismutase (SOD) activity (using colorimetric method) and the expression of Nrf2 and HO-1 protein (by Western blot) in lung tissues and for microscopic examination of pathological changes of the lung which were scored.Results Compared with group TLV,the W/D ratio,pathological scores,MDA content,and expression of Nrf2 and HO-1 were significantly increased,and the SOD activity and oxygenation index at T2,3 were decreased in OLV and Cur groups (P ＜ 0.05).Compared with group OLV,the W/D ratio,pathological scores,and MDA content were significantly decreased,and the SOD activity,oxygenation index at T3,and expression of Nrf2 and HO-1 were increased in group Cur (P ＜ 0.05).Conclusion Curcumin pretreatment reduces ventilator-induced lung injury through promoting the expression of Nrf2 protein in lung tissues in rabbits.