Objective To investigate the factors affecting the protective effects of morphine preconditioning on murine hippocampal neurons against anoxia/reoxygenation (A/R) injury and the underlying mechanisms.Methods Hippocampal slices (400 μm thick) were prepared using hippocampi isolated from decapitated mice. A/R injury was simulated in vitro using artificial cerebral spinal fluid (ACSF) deprived of O_2 and glucose for 20 min followed by reoxygenation and glucose supply for 2 h. The experiment was performed in 4 parts: I .The slices were incubated with 5 different concentrations of morphine (0.1, 0.3, 0.5, 1.0, 3.0, 10.0 /μmol/L) for 30 min at 30 min before A/R; Ⅱ. The slices were incubated with morphine 3.0 /μmol/L for 5 different periods of time (5, 15, 30, 45, 60 min) at 30 min before A/R; Ⅲ. The slices were incubated with morphine 3.0 μmol/L for 30 min followed by A/R at 6 different intervals (0, 5, 15,30,60, 120 min); Ⅳ. The slices were incubated with (a) chelerythrine (a non-selective PKC antagonist) 10 /μmol/L or (b) εVl-2 (a selective nPKCe isoform antagonist) 2 μmol/L or (c) AIP 2 μmol/L (a selective CaMK Ⅱ antagonist) or (d) MK-801 10 μmol/L (a non-competitive NMDA receptor blocker) for 30 min and then for another 30 min together with morphine 3.0 μmol/L before A/R at 30 min interval. The survival rates of the hippocampal neurons were assessed by TTC staining. Results Neuronal survival rates were significantly higher in morphine preconditioning groups which preconditioned with morphine (0.5-10.0 μmol/L) for 15-60 min at an interval of 0-60 min before A/R than in A/R group. Increase in neuronal survival rate induced by morphine preconditioning was partially blocked by chelerythrine or εV1-2 or AIP or MK-801. Conclusion Preconditioning with appropriate concentrations of morphine (0.5-10.0 μmol/L) for appropriate period of time (15-60 min) at appropriate interval (within 60 min) before A/R can protect hippocampal neurons against A/R injury through activation of nPKCε, NMDA receptor and CaMKⅡ.

Objective:To investigate the changes and significances of serum cystatin C and transforming growth factor-β1 levels for the neonatal asphyxia.Methods:Forty-six asphyxia newborns were chosen as the asphyxia group,and thirty healthy newborns were selected as the control group.The TGF-β1,CysC,BUN,Scr,and GFR levels of both groups were detected on the 1st,3rd,7th day after hospitalization.According to the renal injury,the 46 newborns were divided into normal group and asphyxia group,and the serum indexes were detected and analyzed.Results:On the 1st,3rd,7th day after hospitalization,the TGF-β1,GFR of asphyxia group was obviously increased and was lower than those of the control group (P＜0.05);the level of CysC,BUN,Scr in both groups were decreased,and the change degree in asphyxia group were higher than that of the control group (P＜0.05);the CysC,BUN,Scr in renal injured group were higher than those of normal group,and TGF-β1,GFR were much lower (P＜0.05).Additionally,TGF-β1 level of renal injured group was negatively correlated to the BUN and Scr,and positively correlated with the GFR (P＜0.05).The level of serum CysC in renal injured group was positively correlated to BUN and Scr and negatively correlated to GFR (P＜0.05).Conclusion:The serum TGF-β1,CysC in asphyxia newborns had significant changes compared with the healthy newborns and was correlated to the renal injured indexes,which had clinical directive significance on the early diagnosis,condition judgment,and prognosis of neonatal asphyxia with renal injury.

Objective To analyze the clinical data of patients with hilar cholangiocarcinoma (HCCA),and to compare the therapeutic effects of different methods on treating these patients.Methods The clinical data of 101 patients with HCCA in China-Japan Union Hospital of Jilin University were analyzed.Results The overall 1-year and 2-year survival rates in the radical operation group were 95.5％ and 40.9％,respectively.There was a significant difference between the radical operation group and the palliative resection group (P ＜ 0.05).The overall 1-year and 2-year survival rates in the palliative resection group were 75.0％ and 16.7％,respectively,which were much better than those in patients treated with PTCD,biliary stent on open abdominal biliary drainage (P ＜ 0.05).There were no significant differences among the PTCD,biliary stent and open abdominal biliary drainage groups (P ＞ 0.05).Conclusions Radical HCCA resection is still the best and the first choice treatment for patients with HCCA.The therapeutic effects of radical operation were much better than those of palliative resection,biliary stent,PTCD and open abdominal biliary drainage.

As a basic amino acid, histidine has a pK_a close to the acidity of the tumor microenvironment, thus the charge and solubility of histidine are able to vary as the pH changes. Under a neutral environment, histidine is not charged and exhibits hydrophobic properties, while it can be protonated and becomes hydrophilic when exposed to mildly acidic pH, such as tumor microenvironment. Therefore, histidine is widely used in the design of drug delivery systems to target the mildly acidic pH of tumor microenvironment. This article reviews the recent progresses of histidine-based tumor-targeting drug delivery systems, and summarizes the principles on promoting internalization and tuning drug release by taking advantage of histidine. Finally, we point out the common issues on histidine application and illustrate its future prospects.

This study aims to explore the improvement and the mechanism of the Alisma plantago-aquatica Linn. (ApL) on chronic glomerulonephritis (CGN). All animal experiments were followed the regulation of the Experimental Animal Ethical Committee of Shanghai University of Traditional Chinese Medicine. CGN mouse model was established by a single tail-vein injection of doxorubicin (Dox) (20 mg·kg^-1). One week after Dox administration, the mice received water extract of ApL (85 and 255 mg·kg^-1) by gavage once a day for 14 days. At the end of experiment, the urine albumin-to-creatinine ratio (ACR), serum albumin (ALB), blood urea nitrogen (BUN) and serum creatinine (SCr) were detected, kidney histopathological H&E staining was analyzed. Active ingredients and action targets of ApL were collected from TCMSP database, and CGN-related targets were obtained from Genecards database. STRING platform was employed to perform protein-protein interaction (PPI), and Metascape platform was used for KEGG pathway and GO enrichment analysis. The results of experiments demonstrated that ApL (85 and 255 mg·kg^-1) could reduce the ACR and the content of SCr and BUN, and increase the content of ALB in mice. Network pharmacology results predicted that nuclear factor kappa-B (NF-κB)-related pathway and biological process of oxidoreductase activity regulation may be involved in the ApL-provided amelioration on CGN. The verification results showed that ApL could inhibit the activation of NF-κB and the expression of inflammatory factors in mice, and reduce the activity of renal myeloperoxidase (MPO). Meanwhile, ApL promoted the activation of nuclear factor erythroid 2-related factor 2 (Nrf2) and increased the expression of its downstream gene mRNA, and reduced the level of renal malondialdehyde (MDA) and reactive oxygen species (ROS), and further elevated renal glutathione (GSH) level. Based on network pharmacology combined experiments, this study found that ApL may improve CGN in mice through multiple targets and multiple pathways, in which the inhibition of NF-κB signaling and the activation of Nrf2 signaling may be important mechanisms involved.

Objective To observe the changes in transfer rates of epigoitrin, forsythoside A and forsythin in Ganmao Tuire Granules prescription medicinal materials by water decoction extraction, vacuum concentration and atmospheric concentration.Methods Prescription medicinal materials fromGanmao Tuire Granules were decocted in water and the decoction was condensed by vacuum and atmospheric concentration process. The contents of epigoitrin, forsythoside A and forsythin at different states were measured by HPLC for transfer rates.Results The transfer rates of epigoitrin, forsythoside A and forsythin were 72.97%, 29.27%, 72.84%, respectively after decocted in water. The RSD of epigoitrin, forsythoside A and forsythin were less than 2% in the process of vacuum concentration. The transfer rates of epigoitrin, forsythoside A and forsythin by atmospheric concentration decreased by 13.51%, 7.38% and 4.24% respectively compared with vacuum concentration when concentrated to the same state.Conclusion The method for simultaneous determination of epigoitrin, forsythoside A and forsythin by HPLC has been established. A more suitable extraction and concentration process should be chosen for the preparation ofGanmao Tuire Granules.

Objective To investigate the impact of AG490 on the blood-brain barrier (BBB ) permeability and the expression of interleukin-6 (IL-6 )and tumor necrosis factor-α(TNF-α)after traumatic brain injury (TBI)in rats. Methods A total of 144 healthy male SD rats were randomly divided into a control group,a trauma group,and an AG490 intervention group (n=48 in each group). The rats in each group were redivided into four subgroups (4 h,1 d,3 d,and 7 d subgroups)according to the time points after cerebral injury (n=12 in each subgroup). A brain trauma models were induced by hydraulic shock method. Evans blue was used to determine the changes of the BBB permeability after cerebral injury in each group. Real-time fluorescence quantitative PCR was to detect the expression levels of TNF-αand IL-6 mRNA in rat brain tissue. Immunohistochemistry was used to detect the expression of human phospho tyrosine kinase (P-JAK2). Results (1)The permeability of BBB:The permeability of BBB increased at 4 h,1 d,3 d and 7 d after brain injury in the trauma group (Evans blue permeation:10. 4 ± 1. 2,16. 0 ± 1. 4,22. 3 ± 2. 0,and 8. 4 ± 0. 9μg/g,respectively). Compared with the control group, there were significant differences (all P<0. 01). The Evans blue permeation of the AG490 intervention group were 9. 1 ± 1. 0,12. 8 ± 1. 1,17. 5 ± 1. 4 and 7. 1 ± 0. 8μg/g,respectively at each time point,and they were all significantly lower than those of the trauma group (all P<0. 01). (2)The expression of IL-6 and TNF-α mRNA:The expression levels of IL-6 mRNA and TNF-α mRNA at 4 h,1 d,3 d and 7 d after traumatic brain injury in the trauma group were 2. 31 ± 0. 35,2. 73 ± 0. 35,3. 32 ± 0. 29,2. 14 ± 0. 24 and 7. 46 ± 1. 18,9. 42 ± 1. 54,13. 76 ± 1. 89,and 6. 28 ± 1. 00,respectively,they were all significantly higher than those of the control group (all P<0. 01). The expression levels of IL-6 mRNA and TNF-α mRNA of the AG490 intervention group were 1. 14 ± 0. 22,1. 54 ± 0. 23,1. 94 ± 0. 32,1. 26 ± 0. 21 and 5. 57 ± 0. 88, 7. 78 ± 1. 02,11. 51 ± 1. 29,and 5. 05 ± 0. 97,respectively,they were all lower than those of the trauma group,but they still higher than the control group. There were significant differences (all P<0. 01). (3 )The expression of P-JAK2:The expression levels of P-JAK2-positive cells at each time point after traumatic brain injury in the trauma group were significantly higher than the control group (all P<0. 01),they were 17. 4 ± 2. 7,56. 2 ± 6. 7,26. 1 ± 5. 4,and 15. 3 ± 2. 5,respectively;those of the AG490 intervention group were 12. 2 ± 1. 4,41. 5 ± 4. 6,19. 4 ± 4. 1,and 9. 6 ± 2. 0,respectively,they were all lower than those of the trauma group,but still higher than the control group. There were significant differences (all P<0. 01). Conclusion During the acute phase after TBI,AG490 may activate the factor signaling pathways by inhibiting the non-receptor tyrosine kinase/signal transduction and transcription,significantly inhibit the expression of brain tissue inflammatory cytokines IL-6 IL-6 and TNF-α,reduce the BBB damage,and help to reduce secondary brain injury.

Objective To express EV71 VP1 in prokaryotic expression system,initially evaluate the ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody.Methods Construct the prokaryotic expression plasmid pET30a (+)-VP1.Induced expression in Transetta (DE3) with IPTG and identified by Western blot.After purified with HisBind Protein Purification Kit,its ability of blocking EV71 infection and the neutralizing activity of its polyclonal antibody were analyzed.Results Plasmid PET-30a(+)-VPI was constructed successfully and the objective protein was expressed effectively.The ELISA titer of the polyclonal antibody was 1:3200 while neutralizing titer was 1:16 and the recombination protein lost the ability of blocking EV71 infection.Conclusion The recombination protein can stimulate mice to produce antibodies and the polyclonal antibody shew neutralizing activity but the recombination protein lost the binding activity to receptors probably due to the wrong advanced structure.

Objective To study the anti-oxidative stress effects of benazepril and candesartan.Methods SHRs of 12 weeks old were given benazepril(10 mg/kg?d,n=9)or candesartan(4 mg/kg?d,n=9)or combina- tion(Ben:10 mg/kg?d+Can:4 mg/kg?d)for 12 weeks.The tail arterial pressure was measured every two weeks.At end of study,pathological changes in the thoracic aorta,activity of SOD,serum contents of NO and hydroxy radicals,plasma Ang Ⅱ and cGMP,eNOS and P22~(phox)protein expressions in aortic tunica intima were de- termined.Results The thoracic aorta wall was thickened markedly in SHRs,and blood pressure,hydroxy radi- cal,Ang Ⅱ and P22~(phox)protein expression were increased significantly,while the serum NO,level of cGMP and eNOS expression were decreased.Benazepril(Ben)or Candesartan(Can)inhibit the thickening of vessel wall, enhance the activity of SOD(Ben:68.7?2.1,Can:65.6?4.2 vs SHR:48.8?3.2 U/mL,P

Objective To explore the application of Oxybuprocaine Hydrochloride Gel in colonoscopy examination. Methods 1000 patients received colonoscopy examination were randomly divided into experimental group and control group, 500 cases in each. Oxybuprocaine Hydrochloride Gel was applied on anal region in the experimental group before endoscopy for perianal anesthesia and lubrication. Paraffin oil was used in the control group to lubricate perianal and enteroscopy. The success rate of primary insertion, visual analogue pain score (VAS) score, examination time and postoperative complications were compared between the two groups. Results In the experimental group, the success rate of primary insertion (95%) was higher than that of the control group (76%), and the pain score was lower than that of the control group.The examination time was shortened, and the difference was statistically significant (P < 0.05). Conclusions Oxybuprocaine Hydrochloride Gel applied to colonoscopy can effectively reduce the patient's pain and discomfort, improve the success rate of primary insertion, shorten the examination time. And the method is worthy of clinical popularization and application.