Objective To observe and explore the effect and clinical value of percutaneous electrical stimulation on nerve regeneration after end-to-side neurorrhaphy in rats. Methods Thirty-two SPF male S-D rats were randomly divided into four groups ( n = 8 ): group A, the normal control group; group B, with end to end neurorrhaphy of musculocutaneous nerve injury matched to the ulnar nerve; group C, with end to side neurorrhaphy of musculocutaneous nerve injury matched to the ulnar group; and group D, with end to side neurorrhaphy of musculocutaneous nerve injury matched to the ulnar nerve plus postoperative transcutaneous electrical stimulation ( 30 min per day for 6 weeks ) . Electromyography, postoperational nerve conduction velocity, the histological and ultrastructural changes of the nerve fibers were examined, and NF-200 expression in frozen sections was observed using imunohistological staining, to assess the recovery of muscle strength of the diseased side limb and the neuroregeneration in the rats after treatment. Results The amplitude and conduction velocity of the groups C and D were lower than that of the group A, the latency was higher than that of the group A, while the amplitude and conduction velocity of the group D were lower than that of the group C,and the latency was higher than that of the group C. The wet weight ratio of the biceps brachii muscle and the cross-sectional area of muscle fibers in the groups B, C and D were lower than those in the group A, and the recovery of muscle in the group C was the worst. The expression of NF-200 in the rats of groups B, C and D was significantly lower than that in the group A, and the expression of NF-200 in the group D was significantly higher than that in the group C, but still significantly less than that in the group B ( P < 0. 05 ) . Electron microscopy showed mature myelinated fibers in the group B, whereas unmyelinated fibers were the main component and the myelin sheath was poorly developed in the group C. The myelin regeneration in the group D was better than that in the group C, but still some unmyelinated nerve fibers were seen. Conclusions The percutaneous electrical stimulation can effectively promote nerve axonal regeneration and can delay the atrophy of the target muscle after end-to-side neurorrhaphy. Though there is difference compared with the end-to-end neurorrhaphy, the end-to-side neurorrhaphy is still an effective method in clinical repair of peripheral nerve injury.

Objective To explore the effect of intermedin ( IMD ) and adrenomedullin ( ADM ) on cerebral microcirculation in rats with cerebral ischemia. Methods Rat cerebral ischemia ( CI) model was established by middle cerebral artery occlusion. 40 SPF male adult Sprague-Dawley ( SD) rats were randomly divided into three groups:CI+NS ( normal saline) group, CI+ADM group and CI+IMD group, which were used to observe the changes of brain surface microcirculatory perfusion with a laser Doppler flowmeter. Results The differences of brain surface microcirculatory perfusion were statistically significant among the CI+NS group, CI+ADM group and CI+IMD group ( F=53. 426, P<0. 05 ) . Multiple comparison showed that the brain surface microcirculatory perfusion in the CI+IMD group was higher than that of the CI+NS group and CI+ADM group. Conclusions Intermedin can improve the cerebral microcirculation in rats with cerebral ischemia, and its therapeutic effect is better than adrenomedullin.

Objective To investigate the effect of hypobaric hypoxia and cold exposure on brown adipose tissue in mice. Methods Twenty-four 6-week old SPF C57BL/6 male mice were randomly divided into 4 groups with 6 mice in each group: normal atmospheric pressure and temperature group ( 18~22℃, 20~60 m ) ( NTNP ) , low atmospheric pressure and normal temperature group ( 18~22℃, altitude of 5000 m ) ( NTLP ) , normal atmospheric pressure and cold exposure group(0~6℃, altitude of 20 ~60 m)(LTNP), low atmospheric pressure and cold exposure group(0 ~6℃, altitude of 5,000 m)(LTLP). The experimental period was 4 weeks. The body weight was measured at the beginning and end of the experiment. By the end of the four-week trial, the back and inguinal fat were dissected and observed by histology using HE staining. The expression of UCP-1 as the marker of brown adipose tissue in the back fat was detected by qPCR and western blot. Results The body weight gain of NTNP group was higher ( P< 0. 05 ) than the other three groups. Meanwhile, the color of the back and groin fat tissue of mice of LTNP and LTLP groups were darker, the blood supply in mice of these two groups was richer than the NTLP group. The volume of adipose tissue of NTNP group was higher than others. The histology showed that the back adipose cells of the mice were smaller and darker and full of multilocular lipid droplets, exhibiting a typical morphology of brown fat cells. Compared with the NTNP and NTLP groups, the mRNA and protein levels of UCP-1 were higher under cold exposure, while low atmospheric pressure had a tendency to reduce the mRNA expression of UCP-1. Conclusions The formation of brown fat is affected by the imitated conditions of low atmospheric pressure and cold exposure, and is more closely related to the decresed temperature.

Objective To explore the effects of Toddalia asiatica extract( TAE) on oxidative damages in mice with myocardial ischemia and hyperlipidemia. Methods Fifty SPF male KM mice were randomly divided into 5 groups:control group,model group,simvastatin group (5 mg/kg),high-dose TAE group (400 mg/kg) and low-dose TAE group (100 mg/kg) . Hyperlipidemia was induced by feeding milk for 4 weeks. At the same time,the corresponding drug was given by oral administration with 20 mL/kg for 28 d. At the end of four-week treatment, isoprenaline hydrochloride ( Iso ) was subcutaneously injected every 24 hours for three times. The body weight, electrocardiogram, heart and liver indexes, myocardial water content, and the serum levels of TC, TG, HDL, LDL, SOD, MDA and GPx were measured. Results TAE significantly improved the electrocardiogram changes induced by Iso and milk in the mice, weight lose, inhibited the heart and liver indexes, reduced the myocardial water content, decreased the levels of TC, TG, LDL and MDA, and increased the contents of HDL, SOD and GPx. Conclusions TAE may play an important role in cardiovascular protection by regulating oxygen free radical metabolism and improving oxidative damages.

Objective To explore the effect of sitostero-3-O-glucoside on proliferation, apoptosis and collagen synthesis of keloid fibroblasts ( KF) . Methods Cell viability was measured by MTT assay after cells were cultured with 0 (control), 3. 125, 6. 25, 12. 5, 25, 50, 100 μg/mL sitostero-3-O-glucoside. Cell proliferation was measured by EdU staining. Cell apoptotic rate and cell cycle were measured by flow cytometry. The level of COL I and COL III was detected by enzyme linked immunosorbent assay (ELISA). Results 3. 125, 6. 25, 12. 5, 25, 50, 100 μg/mL sitostero-3-O-glucoside reduced cell viability ( P< 0. 01 ) , increased inhibitory rate of cell proliferation ( P< 0. 01 ) , IC50 =27. 54 ± 2. 18 μg/mL. Compared with the control group, 12. 5, 25, 50μg/mL sitostero-3-O-glucoside reduced the cell proliferation rate (P< 0. 01), increased the cell early and late apoptotic rates (P< 0. 01), the cell cycle was arrested at G1 phase (P< 0. 01), and the levels of COL I and COL III were down-regulated (P< 0. 01). Conclusions The results show that sitostero-3-O-glucoside can significantly inhibit the proliferation, induce cell apoptosis and inhibit the synthesis of collagen of keloid fibroblasts.

Objective To observe the effect of different storage time on 14 blood biochemical indexes in rats. Methods Randomly selected 40 adult SD rats were included in this study. Fasting venous blood samples were collected, serum was separated, sealed, and stored in the refrigerator (4℃ and -20℃). The serum parameters were detected at 0 h,4 h,24 h,96 h and 7 d, respectively, using an automatic biochemical analyzer. A total of 14 blood biochemical indexes were detected, including alanine aminotransferase ( ALT ) , aspartate aminotransferase ( AST ) , alkaline phosphatase (ALP), total protein (TP), albumin (ALB), creatinine (CREA-J), uric acid (UA), urea nitrogen (UREA), blood glucose ( GLU) , total cholesterol ( TC) , triglyceride ( TG) , low density lipoprotein cholesterol ( LDL-C) , creatine kinase ( CK) and lactate dehydrogenase ( LDH) . The effects of serum storage time on blood biochemical results were compared. Results The trends of blood biochemical data in male and female rats were consistent. C ompared with the indexes of serum preserved at 4℃ for 0 h, the ALP was significantly reduced after storage for 4 h, 24 h, 96 h, and 7 d (P< 0. 05), ALB were significantly increased after 96 h and 7 d (P< 0. 01), CREA-J was significantly increased after 96 h, 7 d (P<0. 05), UA was significantly increased after 24 h, 9 h, and 7 d (P < 0. 01), and no significant changes in other indicators ( P> 0. 05 ) . Compared with the values of 0 h serum, the serum preserved at -20℃ showed that ALT was significantly increased after 7 d (P < 0. 01), AST significantly increased after 96 h and 7 d (P< 0. 05), TP significantly decreased after 4 h and 24 h ( P< 0. 05 ) , ALB significantly increased after 4 h, 24 h, 96 h, and 7 d ( P< 0. 01 ) , CREA-J significantly increased after 24 h, 96 h, and 7 d (P< 0. 01), UA significantly increased after 4 h, 24 h, 96 h, and 7 d (P< 0. 01), TC significantly increased after 4 h, 24 h, 96 h, and 7 d (P< 0. 01), TG significantly increased after 96 h and 7 d (P< 0. 05), CK significantly increased after 96 h and 7 d (P< 0. 05), LDH significantly increased after 96 h and 7 d ( P < 0. 05 ) , and no significant changes in other indicators ( P > 0. 05 ) . Conclusions The biochemical tests of rat serum should be immediately performed as they were collected, especially for ALP test. For the sera stored at 4℃, the test should be finished in different times:UA test in 4 hours, ALB and CREA-J test in 24 hours, and ALT, AST, TP, UREA, GLU, TC, TG, LDL-C, CK, and LDH test in 7 days.

Objective To analyze the electrocardiography ( ECG ) data of pressure overload ?induced chronic heart failure rats. Methods Totally 20 SD rats were randomly divided into sham operation group and heart failure group. Heart failure rats were induced by abdominal aorta constriction. Echocardiogram measurement demonstrated the occurrence of cardiac function. Two lead ECG parameters of limb a was measured and statistically analyzed. Results Ten weeks after operation, there was a increase in heart rate, P amplitude, P duration and R amplitude comparing by those of the sham operation group (P < 0?05). ECG showed a significant and ubiquitous J point elevation (P < 0?05), with ST segment notable depression ( P < 0?05 ) . Conclusions ECG in pressure?overload chronicity heart failure rats exhibits obviously characteristic features. ECG is an useful tool for objective and accurate assessment of cardiac function in rats.

Objective To determine if aquaporin1 ( AQP1) and aquaporin5 ( AQP5) are expressed in the alveolar-capillary membrane in rats, and to investigate the changes of AQP1 and AQP5 expression in the rat with acute lung injury.Methods The distribution of AQP1 and AQP5 in alveolar capillary membrane was investigated by immunohistochemistry and immunoelectron microscopy with affinity-purified antibodies to human AQP1 and AQP5.The possibility that alveolar capillary membrane AQP1 and AQP5 undergo altered regulation was studied by a rat model established using intra-tracheal instillation of lipopolysaccharide (LPS).Results Immunolabelling showed that AQP1 was stained primarily in the microvascular endothelium of normal lungs, while AQP5 was expressed in type I pneumocytes. Immunohistochemical analysis showed a significant decrease in the expression of AQP1 and AQP5 in injured lungs at 4 -48 h after LPS instillation.AQP1 protein was resumed partly at 24 h after LPS instillation and steroid administration, whereas AQP5 was unchanged.Conclusions The decreased expressions of AQP1 and AQP5 in injured lungs suggest that both of them may play a role in abnormal fluid transportation.

Objective To investigate the method to prevent the displacement of tissue expander implanted between the panniculus carnosus and deep fasia in rabbits .Method 40 rabbits were divided into two groups randomly: the experimental group and the control group .Through an incision on the back , a pocket was made by dissecting between the rabbit panniculus carnosus and deep fasia .The panniculus carnosus at the pocket boundary were sutured with the deep fascia inside the pocket by interrupted 3-0 sutures.A 50 mL kidney-shaped tissue expander was implanted .The expander was inflated with 10 mL saline after the wound was sutured .7 days after implantation , black tattoos were made on the skin around the expander to mark the site of expander .Two months after implantation , the expanders in the experimental group were inflated with 10 mL saline weekly to a total of 140 mL.7 days after implantation the expanders of control group were inflated with 10 mL saline weekly to a total of 140 mL.The incidence of expander displacement , dehiscence of incision , reversal of injection pot and death were recorded .Results There was no expander displacement in the 19 rabbits of experimental group .All the expanders were located within the tattoo area .There was no expander displacement in 11 rabbits of the control group .4 expanders of the control group moved from the back to belly .The difference between them was statistically significant ( P <0.05 ) .The incidence of dehiscence of incision in the experimental group was not significantly lower than that in the control group ( P >0.05 ) .Meanwhile there was no significant difference in the incidences of reversal of injection pot , infections and deaths ( P >0.05 for all ) .Conclusions Sutures between the panniculus carnosus and the deep fascia at the pocket boundary asisted by delayed inflating can prevent the displacement of tissue expander implanted between the panniculus carnosus and deep fasia in rabbits , thus to obtain effective expansion .

Objective To explore different methods for establishing animal models of Ureaplasma urealyticum ( UU) infection and to select the best method of establishment of this disease .Methods SPF female BALB/c mice and Wistar rats were multiple-infected with small dose of UU liquid or single-infected with high-dose UU liquid, pretreated with estradiol benzoate, to establish models of Ureaplasma urealyticum infection.Cervical secretion was taken for UU culture at 14, 28, 42 and 56 days after the first time UU treatment , and the reproductive organs were taken for histopathological examination.Results Gross examination of the UU-infected rats showed generally less serious lesions than the mice .The low-dose estrogen group was worst , showing hyperemia and edema , cervical hypertrophy , hydrosalpinx , enlargement , and more rigid with poor elasticity .Significant differences were observed in the vaginal UU colonization rates between the low -dose estrogen and low-dose non-estrogen groups , high-dose estrogen and non-estrogen groups , low-dose estrogen and high-dose estrogen groups, low-dose estrogen and non-estrogen groups, in both the mice and rats (P<0.05 for all).The same dose estrogen treatment caused a higher UU colonization rate in the mice than rats ( P<0.05 ) .Histological examination showed that the low-dose estrogen treated rats had no obvious pathological changes , except loose tissue edema in some rats , but apparent pathological changes were observed in the low-dose estrogen treated mice .Conclusions The vulvar lesions in the mice are worse than in the rats .Estrogen pre-treatment can increase the Ureaplasma urealyticum colonization rate in the animals.The UU colonization rate in the vagina of mice is higher than in the rats , and multiple low-dose UU treatment produces a higher UU colonization rate than a single high-dose UU treatment.Therefore, BALB/c mice are more susceptible to Ureaplasma urealyticum infection, with more serious pathological changes , and are more suitable for experimental studies of related diseases .