Objective To evaluate the effects of sevoflurane preconditioning on wnt/glycogen synthase kinase-3 beta (GSK-3β)/β-catenin signaling pathway during myocardial ischemia-reperfusion (I/R) injury in rats in vitro.Methods Ault male Wistar rats,weighing 220-280 g,were heparinized and anesthetized with intraperitoneal 3％ pentobarbital 30 mg/kg.Their hearts were rapidly excised and perfused in a langendorff apparatus with oxygenated (95％ O2-5％ CO2) K-H solution at 37 ℃.After 15 min of equilibration,36 isolated hearts were randomly divided into 3 groups (n=12 each) using a random number table:sham operation group (group S),group I/R and sevoflurane preconditioning group (group SP).After 30 min of equilibration,the hearts were continuously perfused for 150 min in group S.The isolated hearts were subjected to 30 min of ischemia followed by 120 min of reperfusion.In SP group,the hearts were perfused for 15 min with K-H solution containing 2.4％ sevoflurane,followed by 5 min washout before reperfusion.At the end of equilibration and 30 min of reperfusion,HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP) and ± dp/dtmax were recorded.The severity of arrhythmias was assessed during reperfusion.At 60 min of reperfusion,3 hearts in each group were chosen for measurement of expression of wnt3a,phosphor-GSK-3β (p-GSK-3β) and β-catenin (by Western blot).At 120 min of reperfusion,6 hearts in each group were chosen for determination of myocardial infarct size by TTC staining.Results Compared with group S,HR,LVDP,+dp/dtmax and -dp/dtmax were significantly decreased,and LVEDP was increased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were increased,and the expression of wnt3a,p-GSK-3β and β-catenin was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+dp/dtmax and-dp/dtmax were significantly increased,and LVEDP was decreased at 30 min of reperfusion,arrhythmia scores and the percentage of myocardial infarct size were decreased,and the expression of wnt3a,p-GSK-3β and β-catenin was up-regulated in group SP.Conclusion Sevoflurane preconditioning attenuates myocardial I/R injury by activating wnt/GSK-3β/β-catenin signaling pathway in isolated rat hearts.

Objective To evaluate the effects of sevoflurane preconditioning on zonula occludens-1 (ZO-1) during myocardial ischemia-reperfusion (I/R) in rats in vitro.Methods Adult male Wistar rats,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital 30 mg/kg and heparinized.Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％ CO2 at 37 ℃.Eighteen isolated rat hearts were randomly assigned into 3 groups (n =6 each) using a random number table:control group (group C),group I/R and sevoflurane preconditioning group (group S).At 10 min of equilibration,the hearts were perfused with K-H solution for 110 min in group C,the hearts were perfused with K-H solution for 20 min,and then subjected to 30 min of ischemia followed by 60 min of reperfusion in group I/R,and the hearts were perfused with K-H solution saturated with 3％ sevoflurane for 15 min followed by 5 min washout,and then subjected to 30 min of ischemia followed by 60 min of reperfusion in group S.At the end of equilibration,immediately before ischemia,and at 15 and 60 min of reperfusion (T1,2),HR,left ventricular end-diastolic pressure (LVEDP),left ventricular developed pressure (LVDP),+ dp/dtmax and-dp/dtmax were recorded.The development of arrhythmias was recorded during reperfusion and scored.At 60 min of reperfusion,myocardial specimens were obtained from the apex of heart for determination of the expression of ZO-1 in myocardial tissues (by Western blot) and for observation of distribution of ZO-1 and connexin43 (Cx43) (by immunofluorescence).Results Compared with group C,HR,LVDP,+ dp/dtmax and-dp/dtmax were significantly decreased and LVEDP was increased at 15 and 60 min of reperfusion,scores of arrhythmia was increased,and ZO-1 expression was down-regulated in I/R group.Compared with group I/R,HR,LVDP,+ dp/dtmax and-dp/dtmax were significantly increased and LVEDP was decreased at 15 and 60 min of reperfusion,arrhythmia was decreased,and ZO-1 expression was up-regulated in group S.ZO-1 and Cx43 were co-localized at the intercalated disk.ZO-1 was redistributed in the lateralization of the membrane and co-localized with Cx43 in group I/R.The incidence of ZO-1 lateralization was significantly decreased in group S.Conclusion The mechanism by which sevoflurane preconditioning decreases reperfusion arrhythmia is related to inhibition of down-regulation of expression and redistribution of ZO-1 and inhibition of redistribution of Cx43 in rats.

Objective To explore the optimal dose and ways of anesthesia for creating a rat model of spastic paralysis with intermittent bilateral common carotid artery ligation. Methods 60 Wistar rats were randomly divided into groups A, B, C, D, E and F. Group A was anaesthetized with 10% chloral hydrate 5 ml/kg injected subcutaneously, while group B with 4 ml/kg subcutaneously, group C with 4 ml/kg intraperitoneally, group D with 3 ml/kg subcutaneously, group E with 3 ml/kg intraperitoneally, group F with 2 ml/kg subcutaneously. The onset and duration of anesthesia, and intraoperative and postoperative mortality were compared. Results All the rats in the group A died during anesthesia, while the group F did not achieve the depth of anesthesia, and abandoned. The onset time was (6.5±0.7) min, maintained (121.4± 3.9) min, mortality was 0 in the group B, and it was (5.5±1.1) min, (122.0±3.6) min, 30% in the group C; (9.6±0.8) min, (106.7±3.7) min, 0 in the group D, (7.4±1.2) min, (105.3±3.5) min, 20% in the group E. The overall mortality rate was 0 in the groups accepted subcutaneous injected and 25% of intraperitoneal injection. Conclusion Anesthesia with 10% chloral hydrate 4 ml/kg subcutaneous injection is optimal of lower mortality, faster onset and longer maintaining in rats for spastic paralysis model with intermittent bilateral common carotid artery ligation.

Objective To investigate the DNA methylation of skeletal muscle in spastic paralysis rats and correlation with the muscle fiber configuration. Methods 100 5-day old Wistar rats were randomly divided into model group and control group. The former was established the spastic paralysis modle and reared for 30 days. Then, tissues from the gastrocnemius of all the rats were observed with triplicate DNA methylation, myosin heavy chain-I (MHC-I) mRNA with RT-PCR and transmission electron microscopy. Results The DNA methylation was (4.95±0.83)×10% in the model group, significantly less than (6.59±0.75)×10% in the control group (P<0.001); while the MHC-I mRNA was (1.23±0.31), significantly more than (0.44±0.29) in the control group (P<0.001). The Z-line was disordered, and the mitochondria near the Z-line increased, with edema and partially broken in cristae. The balance between the thick and thin filaments was broken, and myofibrils envelope fused. Conclusion Hypomethylation and hyperexpression of MHC-I mRNA have been found in skeletal muscle of spastic paralysis rats, which may result in type I fibers increase. However, there was no sufficient evidence to support the correlation between the DNA methylation and the secondary pathological changes.

Objective:To evaluate the antimicrobial effectiveness of lomefloxacin hydrochloride eye drops from 13 domestic manu-facturers. Methods:The antimicrobial effectiveness of lomefloxacin hydrochloride eye drops was tested based on the antimicrobial ef-fectiveness testing in Chinese Pharmacopoeia(2015 edition,volume 4). Results:The samples from 5 manufacturers could reach level A,those from 3 manufacturers were at level B, and those from 5 manufacturers did not comply with the specification. Conclusion:Some problems exist in the usage of antimicrobial agents in lomefloxacin hydrochloride eye drops from some Chinese manufacturers,and these manufacturers should optimize the type and concentration of antimicrobial agents according to the requirements of Chinese Phar-macopoeia combined with the corresponding prescriptions under the principle of minimum concentration while best effect.

Emerging evidence has demonstrated the vital role of metabolism in various diseases or disorders. Metabolomics provides a comprehensive understanding of metabolism in biological systems. With advanced analytical techniques, metabolomics exhibits unprecedented significant value in basic drug research, including understanding disease mechanisms, identifying drug targets, and elucidating the mode of action of drugs. More importantly, metabolomics greatly accelerates the drug development process by predicting pharmacokinetics, pharmacodynamics, and drug response. In addition, metabolomics facilitates the exploration of drug repurposing and drug-drug interactions, as well as the development of personalized treatment strategies. Here, we briefly review the recent advances in technologies in metabolomics and update our knowledge of the applications of metabolomics in drug research and development.

Objective: To evaluate the therapeutic effect of mouse nerve growth factor (mNGF) combined with conventional rehabilitation training on children with cerebral palsy (CP). Methods: According to the inclusion criteria, 60 children with CP(spastic diplegia) who were treated at the Third Affiliated Hospital of Jiamusi University were selected by double-blind method from June to December in 2016.Sixty children with CP were stratified according to age, gender and Gross Motor Function Classification System (GMFCS) (the subjects were classified into 2 age groups: 0-2 years old and 2-4 years old), and then they were randomly divided by draw method: the control group received routine rehabilitation; the experimental group received mNGF additionally.The children with CP in the experimental group were treated with 18 μg mNGF every day for 10 days and then discontinued for 5 days, with 20 injections per month for 1 course, a total of 3 courses.After 3 courses of treatment, the changes in Gesell Development Diagnosis Schedules (GDDS), Peabody Developmental Motor Scales-Gross Motor (PDMS-GM), Peabody Developmental Motor Scales-Fine Motor (PDMS-FM) and Range of Motion (ROM) of the lower extremity were compared between the 2 groups. Results: After 3 months of treatment, the changes of Gesell(gross motor: 57.40±18.13, 44.87±10.95; fine motor: 64.83±18.04, 62.60±17.34; adaptability: 76.07±14.99, 70.57±11.19; language: 74.20±15.07, 71.23±13.38, personal-social interaction: 67.40±14.10, 61.40±12.96), PDMS-GM(94.33±16.03, 81.13±20.15), PDMS-FM scores(91.53±19.73, 91.10±15.84) and ROM[adductors angle: left (69.67±22.2)°, (49.17±21.82)°; right (69.83±21.63)°, (49.67±21.61)°; popliteal angle: left (160.08±30.02)°, (125.50±25.78)°; right (160.17±22.46)°, (126.00±25.31)°; dorsal flexion angle of foot: left (10.17±6.09)°, (4.33±7.28)°; right (9.83±6.23)°, (4.83±7.48)°] in the experimental group and the control group were all significantly higher than those before treatment, and the differences were statistically significant (all P<0.05). After 3 months of treatment, the Gesell gross motor(57.40±18.13) and PDSM-GM scores(94.33±16.03) in the experimental group were significantly higher than those (44.87±10.95, 81.13±20.15, respectively) in the control group, and the differences were statistically significant (all P<0.05). Conclusions Conventional rehabilitation therapy combined with mNGF has more significant effect on the development and motor function of children with CP than routine rehabilitation training.