Objective To evaluate the effect of ulinastatin on oxidative stress injury to myocardial cells in diabetic rats in vitro.Methods The H9c2 cells were cultured in DMEM culture medium and the cells at the logarithmic growth phase were seeded in 96-well plates (density 1 × 104 cells/ml,200 μl/well) or in 6-well plates (density 1× 105 cells/m1,2 ml/well).The cells were randomly divided into 4 groups (n=18 each) using a random number table:normal control group (group C),high-glucose group (group HG),high-glucose + oxidative stress group (group HG+OS),ulinastatin +high-glucose+oxidative stress group (group U+HG+OS).The cells were cultured in high-glucose DMEM culture medium (25.0 mmol/L) for 48 h in group HG.After the cells were cultured in high-glucose DMEM culture medium for 24 h,H2O2 with the final concentration of 500 μmol/L was added to the high-glucose culture medium,and the cells were continuously cultured for 24 h in HG+OS and U+HG+OS groups.In group U+HG+OS,ulinastatin 400 U/ml was added to the high-glucose culture medium.The cells were collected for determination of cell viability,H9c2 apoptosis,activity of superoxide dismutase (SOD) and contents of malonadehyde (MDA).Apoptosis rate was calculated.The cell culture supernatant was collected for detection of lactate dehydrogenase (LDH) activity.Results Compared with group C,the cell viability and SOD activity were significantly decreased,and the apoptosis rate,MDA content and LDH activity were increased in the other groups.Compared with HG group,the cell viability and SOD activity were significantly decreased,and the apoptosis rate,MDA content and LDH activity were increased in HG+OS and U+HG+OS groups.Compared with group HG+OS,the cell viability and SOD activity were significantly increased,and the apoptosis rate,MDA content and LDH activity were decreased in group U + HG+ OS.Conclusion Ulinastatin can mitigate oxidative stress injury to myocardial cells in diabetic rats,and inhibited cell apoptosis is involved in the mechanism.

OBJECTIVETo investigate the protective effect and mechanism of Xingnaojing(Traditional Chinese Medicine) injection on brain injury in rats.

METHODSSixty-three healthy adult male SD rats were randomly divided into 3 groups (n = 21): sham operation group, model group, xingnaojing group. The model of traumatic brain injury model group and Xingnaojing group used the free fall impact injury method, the sham operation group underwent craniotomy, did not cause brain damage. Xingnaojing group in rats after 10 min by tail vein injection Xingnaojing injection 10 ml/(kg x d), model group and sham operation group were intravenously injected with 0.9% sodium chloride solution, three groups were administered continuously for 7 days. At administration of the seventh days compared the S-100B protein in the serum and neuro specific enolase (NSE) level, the water content of brain tissue, serum superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) content, and neurological function of rats among groups.

RESULTSCompared with the sham operation group, the nerve defect, brain water content, MDA, S100B protein and NSE levels were obvigusly increased in Xingnaojing group and model group; SOD, GSH-Px content decreased significantly; In Xingnaojing group nerve impairment and brain moisture were significantly lower than those of model group, the serum MDA, S-100B protein and NSE levels were significantly lower than those in the model group, the SOD, GSH-Px activity was significantly higher than that in the model group.

CONCLUSIONXingnaojing injection has protective effects on rat brain injury, and its mechanism may be related to reduce brain edema after traumatic brain injury and inhibit the reaction of oxygen free radical, protect nerve cells.

Animals ; Brain Injuries ; metabolism ; prevention & control ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Phosphopyruvate Hydratase ; metabolism ; Rats ; Rats, Sprague-Dawley ; S100 Calcium Binding Protein beta Subunit ; blood ; S100 Proteins ; metabolism ; Superoxide Dismutase ; metabolism

Hydrophidae, one of the precious traditional Chinese medicines, is generally drily preserved to prevent corruption, but it is hard to identify the species of Hydrophidae through the appearance because of the change due to the drying process. The identification through analysis on gene barcode, a new technique in species identification, can avoid the problem. The gene barcodes of the 6 species of Hydrophidae like Lapemis hardwickii were aquired through DNA extraction and gene sequencing. These barcodes were then in sequence alignment and test the identification efficency by BLAST. Our results revealed that the barcode sequences performed high identification efficiency, and had obvious difference between intra- and inter-species. These all indicated that Cyt b DNA barcoding can confirm the Hydrophidae identification.
Animals ; Base Sequence ; China ; Cytochromes b ; genetics ; DNA Barcoding, Taxonomic ; Elapidae ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA

Objective To observe the protective effect of sufentanil pretreatment on the rats with acute gastric mucosa lesion (AGML) induced by water immersion and restrain stress (WIRS) and its effect on TRPV1 mRNA expression in the hypothalamus and gastric mucosa. Methods Thirty male Wistar rats were randomly designed into 3 groups, including the normal control group (Group NC, n = 10), the group treated with WRIS for 6 h (Group WIRS, n = 10) and the group pretreated with sufentanil (Group SF, n = 10). The model of AGML was established by the classic WIRS method , and observed for the general extent of gastric mucosal injury at WIRS for 6 hr, and calculated gastric mucosal injury ulcer index (UI) and the PH value of gastric juice; The quantification of TRPV1 mRNA expression in hypothalamus and gastric mucosa was performed using quantitative real-time PCR; In addition, the activity of super oxide dismutase (SOD) and the level of malondialdehyde (MDA) in serum were detected. Results Compared with group NC, gastric mucosal in Group WIRS was injured more seriously , and the UI and the activity of MDA were also obviously increased , but the change of SOD activity was not apparent; The TRPV1 expression in gastric mucosal decreased apparently. Sufentanil pretreatment could effectively relieve gastric mucosal injury induced by WIRS , and make the UI and the activity of MDA decreased , and up-regulate TRPV1 mRNA expression in the hypothalamus and gastric mucosa. Conclusions Sufentanil pretreatment can effectively relieve AGML induced by WIRS , which may be related to the control of oxidative stress response , the reduced gastric acid secretion , and the upregulation of the TRPV1 mRNA expression in the central and periphera nerve.

Objective To investigate the effect of pretreatment with dexmedetomidine alone or in combination with sufentanil on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Fifty healthy male Sprague-Dawley rats,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg.Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min of reperfusion.The rats were then randomly divided into 5 groups (n =10 each):sham operation group (group S),group I/R,dexmedetomidine pretreatment group (group DP),sufentanil pretreatment group (group SP),and dexmedetomidine + sufentanil pretreatment group (group DS).In group S the anterior descending branch was only exposed but not ligated.Dexmedetomidine 0.5μg/kg and sufentanil 0.1μg/kg were injected intraperitoneally 30 min before ischemia in groups DP and SP,respectively.Dexmedetomidine 0.5 μg/kg and sufentanil 0.1 μg/kg were injected intraperitoneally 30 min bbefore ischemia in group DS.Arterial blood samples were collected at 120 min of reperfusion for determination of serum creatine kinase (CK) and lactic dehydrogenase (LDH)concentrations.The rats were sacrificed at 120 min of reperfusion and hearts were removed for microscopic examination.Myocardial infarct size was calculated.The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in myocardial tissues were measured.Results Compared with group S,the serum CK and LDH concentrations were significantly increased,the myocardial infarct size was enlarged,and SOD activity was decreased in the other groups,MDA content was significantly increased in groups I/R,DP and SP (P ＜ 0.05 or 0.01).Compared with group I/R,the serum CK and LDH concentrations,MDA content and myocardial infarct size were significantly decreased,and SOD activity was increased in groups DP,SP and DS (P ＜ 0.05).Compared with group DS,the serum CK concentration was significantly increased,the myocardial infarct size was enlarged,and MDA content was increased in groups DP and SP,and LDH concentration was significantly increased and SOD activity was decreased in group DP (P ＜ 0.05).The pathological changes were significantly attenuated in groups DP and SP compared with group DS.Conclusion Dexmedetomidine pretreatment can reduce myocardial I/ R injury in rats,dexmedetomidine combined with sufentanil pretreatment provides better efficacy than either alone,and inhibition of lipid peroxidation is involved in the mechanism.

Objective To observe the effects of the preconditioning of ulinastatin on GES-1 cell injury induced by oxygen and glucose deprivation (OGD). Methods GES-1 cells were cultured in vitro and divided into three groups: normal control group (group N), oxygen and glucose deprivation group (group O), and ulinastatin preconditioning group (group U). The OGD model of GES-1 cells were established by glucose-free medium and three-gas incubator for 6h. Ulinastatin was added to group U 12h before the deprivation of oxygen and glucose. The cell viability and apoptosis were determined by cck-8 and flow cytometry respectively. Western Blot was used to examine the protein expression of Caspase-3 and Cleaved Caspase-3. The TRPV1 mRNA expression was measured by quantitative real-time PCR. Results As compared with group N, the viability of GES-1 was decreased, the apoptotic rate and the expression of Caspase-3 and Cleaved Caspase-3 were increased, and the TRPV1 mRNA expression decreased greatly in group O (P < 0.05). As compared with group O, the aforementioned changes were significantly inhibited in group U. Conclusions Ulinastatin preconditioning could effectively inhibit GES-1 cell injury induced by OGD, which may be related to the inhibition of apoptosis and the upregulation of TRPV1 mRNA expression.

AIM: To observe the influence of the selective decontamination of the digestive tract (SDD) and caecosomy/colonic irrigation on gut endotoxin/bacteria translocation following acute severe pancreatitis (ASP). METHODS: Twenty three pigs weighing 16-22 kg were divided into four groups. Group I (n=5): sham-control; Group Ⅱ (n=6): ASP-control; Group Ⅲ (n=6): gntamicin [(8.55?10~5?5.70?10~4) units/time] and nystatin [(1.37?10~5?9.00?10~3) units/time] were fed orally every 8 h for 1 week before the induction of ASP; Group Ⅳ (n=6): caecostomy was performed before the induction of ASP. ASP was induced by infecting 1 mL/kg BW of combined solution of 5% sodium taurocholate and (8-10)?10~6 BAEE units/L of trypsin into pancreas via pancreatic duct. Systemic plasma endotoxin levels were quantified by the chromogenic limulus amebocyte lysate (LAL) technique. Specimens of tissue from mesenteriolum and mesocolon lymph nodes, lung, lymph nodes in hilus pulmonis, pancreas and the samples of both portal and systemic blood were collected before and at 72 h following ASP and cultured for aerobic as well as anaerobic bacteria growth. Positive specimens were subcultured and the bacteria identified by standard procedure. RESULTS: Preventive SDD not only effectively reduced the amount of bacteria in stool (P

To construct a secretory-expression vector of antimicrobial peptide Bactenecin 7(Bac7),and identify the secretory-expression product in L.lactis MG1363 and its bioactivity.The splicing primers of regulation elements and Bac7 gene,which designed according to codon usage preferences of L.lactis MG1363,were chemically synthesized,and the overlap-extension PCR method was used to splice the full length of Bac7 gene.Then the Bac7 gene was linked to expression vector pMG36e to construct pMG36e/Bac7 vector,and pMG36e/Bac7 was transformed into L.lactis MG1363 by electrophoration.RT-PCR and Western blot assays were applied to investigate the expression of the Bac7 gene in L.lactis,and bioactivity of Bac7 in culture supernatant of L.lactis was tested with plate-diffusion method.The results showed that the Bac7 gene and its regulation elements was amplified and cloned in the vector pMG36e successfully,The secretory-expressed Bac7 in L.lactis MG1363 harboring pMG36e/Bac7 was identified by Western blot,and it had high bacteriostatic activity against E.coli.These results indicate that the recombinant L.lactis MG1363 could express bioactive Bac7,which lays a foundation for further study of oral administration of a Bac7-secreting L.lactis to treat intestinal bacteria infection.

Objective:To design Keap1-tat peptide and explore its neuroprotective role on hipocampal CA1 neuron,as well as the effect on spacial learning and memory function following global cerebral ische-mia.Methods:Adult male Sprague Dawley (SD)rats were subjected to global cerebral ischemia (GCI) by four-vessel occlusion for 1 5 min and randomly divided into five groups:sham,sham+Keap1-tat,is-chemia/reperfusion (I/R),Keap1-tat peptide-and vehicle-administrated groups.For Keap1-tat or vehi-cle groups,the rats were treated with Keap1-tat (30,50,1 00 μg in 5 μL 0.9%saline)or the same vo-lume vehicle by intracerebroventricular injection (icv)30 min prior to ischemia.Cresyl violet staining was used to observe the surviving neurons and 4-hydroxy-2-noneal (4-HNE ) and 8-hydroxy-2′-deox-yguanosine (8-OHdG)immunostaining were used to detect the change of markers response to oxidative stress in hippocampal CA1 region.The spatial learning and memory function of the rats was evaluated using Morris water maze.Results:Compared with sham group,the number of surviving neurons in ische-mia-reperfusion and vehicle groups significantly decreased in the hippocampal CA1 region (P<0.05 ), while administration of Keap1-tat significantly decreased the damage following GCI (P<0.05),and the dose of 50 μg existed the most effective neuroprotective role.Furthermore,immunostaining intensity of 4-HNE and 8-OHdG,markers of oxidative stress damage attenuated by Keap1-tat peptide as compared with vehicle group in CA1 region.Of significant interest,the time of finding underwater platform in Keap1-tat group animals was significantly short,and after removing the platform,the probe time of Keap1-tat group animals in the original quadrant where the platform was significantly increased compared with that of vehi-cle and I/R group animals (P<0.05).Conclusion:Keap1-tat peptide can effectively attenuate neuro-nal damage in hippocampal CA1 region and improve learning and memory function,which might bedue to the attenuation of oxidative stress caused by GCI.

Objective To introduce an effective nursing practice for patients with Grave's ophthalmopathy(GO) undergoing endoscopic trans-ethmoid orbital decompression(ETOD). Methods The treatment and nursing experience of 31 cases (57 eyes) undergoing orbital decompression through endoscopic trans-ethmoid approach were reviewed and analyzed. Results Three months after operation, exophthalmos was corrected in all 31 cases (57 eyes) with satisfying result, and the visual acuity was improved in all 9 cases (16 eyes),who complained of visual loss preoperatively. At the 3-months' review, 2 patients presented diplopia on the primary eye position due to newly on-set global displacement. Conclusions Advanced treatment and careful nursing are very important to improve the surgical safety and decrease the complications.