Objective To investigate the effects of methamphetamine (Meth) on the outward K+ currents and elucidate the role of outward K+ channels in Meth induced hippocampal neuron damage.Methods Hippocampal neurons were harvest from 18-day-old embryonic rats and were divided into two groups:the control group and the Meth treated group.Both of 4-AP and TEA sensitive K+ currents were recorded after the treatment of Meth by performing the whole cell patch clamp.Furthermore,the MTT and TUNEL assays were performed to evaluate the effects of K+ channel on hippocampal neuron damage mediated by Meth.For statistical comparison,One-way ANOVA and LSD multiple comparison test or t-test was used.P-value ＜ 0.05 was considered to be statistically significant.Results The density of 4-AP sensitive K+ channel currents in Meth treated group [(120.1 ± 19.6) pA/pF,n =7] were significantly increased when compared with control group [(87.4 ± 12.5) pA/pF,n =10,P ＜0.01] and the increments of the currents induced by Meth was dose dependent.The MTT data showed that the cell viability was obviously decreased in Meth treated group (48.72 ± 4.38) ％ relative to the control group (100.07 ± 3.36) ％.Moreover,application of K+ channel antagonist,4-AP (61.39 ± 3.15)％,and the high K+ solution (78.25 ± 9.42) ％ substantially enhanced the cell viability.The TUNEL assay showed there were protective effects of 4-AP and the high K+ solution against neuron damage observed during cells exposed to Meth.Conclusions The increments of 4-AP sensitive K+ channel currents induced by Meth might be involved in hippocampal neuron damage.

Objective To investigate the relationship of albumin levels with the prognosis and severity of illness in elderly sep-sis patients.Methods This was aretrospective study.108 elderly sepsis patients were enrolled from October 2014 to Decem-ber 2015.All patients were divided into survivors group (83 cases)and death group (25 cases)based on the 28-day progno-sis.The differences of clinical data and laboratory were compared between two groups.According to the albumin levels,all patients were divided into three groups,normal albumin group (≥35 g/L,24 cases),and mild hypoproteinemia group (28 g/L≤ALB<35 g/L,52 cases)and severe hypoproteinemia group (<28 g/L,32 cases),respectively.The mortality rate was compared in the groups.Spearman Correlation Coefficient was used to analyze ALB and other factors.Results In all 108 pa-tients,the 28-day mortality rate was 23.1%,and 77.8% of the patients with hypoproteinemia.ALB level [31.7 (28.3~35.7)g/L vs 25.8 (21.7~31.8)g/L,P<0.001]and Hb [128 (110~140)g/L vs 102 (84~132)g/L,P=0.015]in death group were significant lower than that in survival group.Meanwhile,APACHE II scores [20 (18~23)vs 22 (19~24),P=0.015]and SOFA score [6 (5~6)vs 6 (6~7),P<0.001]were higher than that in survival group.The mortality decreased with the increase of ALB level (43.8% vs 21.2% vs 0%,P<0.05).ALB had a negative correlation with APACHEⅡ score (r=-0.190,P=0.049)and coma (r=-0.311,P=0.001),and had a positive correlation with Hb (r=0.449,P<0.001).Conclusion The incidence of hypoproteinemia was high in elderly sepsis patients.ALB level was associated with prognosis and severity of illness among the patients.

Rat osteoblasts were isolated from the 21-day fetal rat calvarias. The cells were grown in DMEM plus 10% FBS, and were treated for 24 h. With 10 μmol/L TPEN or 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+ . Apoptosis of osteoblasts were measured by flow cytometry, electron microscopy and DNA fragmentation analyzed by gel electrophoresis. In addition, IP3 production and PKC activity were measured in order to show whether they are involved in apoptosis in osteoblast induced by zinc deficiency. The results showed that 10 μmol/L TPEN could induce apoptosis in osteoblast in 24 h. But cells treated with 10 μmol/L TPEN supplemented with 10 μmol/L Zn2+showed no apoptotic changes in 24 h. TPEN significantly reduced the formation of IP3 and PKC activity after 24 h incubation. No differences were observed between the cells treated with TPEN supplemented with Zn2 + simultaneously and the untreated cells. It can be inferred that apoptosis induced by zinc deficiency may be due to the decreased activity of PKC which is impaired by reduced formation of IP3.

Objective To explore the neural damage induced by acute exposure to methamphetamine (METH).Methods The mice were administrated with METH,then the stereotyped behavior of mice was evaluated,and spatial recognition memory was analyzed by Y-maze test.In addition,nitric oxide synthase (NOS) activity was detected by kit,and the apoptotic proteins including Bax,Bcl-2,Caspase-3 were assayed by using Western blot.The DNA injury induced by METH was observed by using the comet assay.Moreover,mitochondrial membrane potential was detected to assess the toxic effects of METH on mitochondria by JC-1.With the Western blot assay,the phosphorylation of MAPK signaling pathways were also investigated.Results Acute METH exposure significantly increased the stereotyped behavior in mice,and spatial recognition ability of mice was obviously decreased.On the molecular level,total nitric oxide synthase (TNOS) and induced nitric oxide synthase (iNOS) were increased,and the apoptotic proteins,such as Bax and cleaved caspase-3 were markedly enhanced.With the comet assay,it showed that METH exposure resulted in DNA damage.In parallel,mitochondrial membrane was damaged which manifested as mitochondrial membrane potential decreased.With the western blot,It was further found that METH enhanced the activation of MAPKs.However,p38 MAPK signahng pathway was demonstrated to be the only one factor involved in METH-induced neural damage.Conclusion METH induced neural damage,and MAPK signaling pathways might be involved in this process,since inhibition of p38 MAPK signaling pathway significantly ameliorated METH-induced neural damage.

Objective To determine whether intermittence irradiation of single blue or white light have an adverse effect on the DNA of newborn infants with hyperbilirubinemia by examining the sister chromatid exchange(SCE)frequency of peripheral blood lymphocytes.Methods The frequency of SCE in lymphocytes of 40 icteric infants treated by different phototherapy(PT) methods was a nalyzed by sister chromatid differetance staining technique (SCD).The patients receiving PT were divided into three groups according to two methods of PT,group A:single blue light,20 cases; group B:single white light,20 cases.Results 1.In group A, there was no difference between the levels of SCE before and after therapy within 3 days;but after 4 days, the levels of SCE increased.2.Obvious changes were observed in group B,and the frequency of SCE increased after 1 day and increased significantly in a dose-dependant manner.3.After treatment, the SCE frequency of group B was higher than that of group A.Conclusions PT has mutagenic effect on newborns with hyperbilirubinemia. The effect of single white light on peripheral blood lymphocytes of neonates is more significant.

Objective To evaluate the clinical efficacy of interventional occlusion of ventricular septal rupture (VSR) complicating acute myocardial infarction (AMI). Methods Six patients with VSR complicated after AMI underwent transcatheter interventional occlusion in Chinese PLA General Hospital between May 2009 to May 2015 were retrospectively analyzed. Their clinical features, interventional treatment protocols, and occurance of postoperative complications were studied. Results Among the 6 patients, VSR were successfully occluded in 5 patients. One patient failed the operation due to instability of occluder after it was deployed and the occluder was retrieved. The paitents died of heart failure 6 months later. Among the 5 patients with successful closure, 4 patients presented mild residual shunt after occlusion and acute left heart failure occurred in 1 patient after operation. Two patients died during hospital stay after operation. Between them, one patient died 3 hours after operation because of cardiac tamponade and the other patient died of cardiogenic shock after withdrawal from IABP. Three patients were followed up until now and follow up echocardiography showed satisfactory cardiac function without heart failure. Conclusions Transcatheter occlusion was a reliable therapy for patients with ventricular septal rupture complicated in acute myocardial infarction which could improve cardiac function and reduce mortality.

Objective To explore the endoscopic features and careinoembryonic antigen (CEA) ex-pressions of colorectal serrated adenomas (SA). Methods From June 2005 to July 2008, 27 patients with colorectal polyps and 26 cases of advanced colorectal cancer (ACC) were enrolled in the study. The pit pat-tern of the suspected lesions were observed with 0. 4% indigo carmine sodium stain and classified according to Kudo classification. The polyps were removed by biopsy, high-frequency electrical excision or endoscopic mucosa resection and the samples of ACC were collected with biopsy forceps. All specimens underwent rou-tine pathological examination and CEA expression was detected by immunohistochemistry. Results There were 47 SAs and 27 other types of polyps in polyp group. Most SAs located in left-side colon and were char-acterized by the bulge semipedunculation. The diameters of 45 serrated adenomas ranged from 3 to 8ram, and the maximal diameter of other 2 were more than 10ram. The most common type of pit pattern in SA was mixed type Ⅱ and ⅢL, with type Ⅱ predominant in a certain percentage. Expression of CEA in SA glands was significantly higher than that in proliferative polyps (P < 0. O1). Candnsion SA is an unique type of colorectal adenoma, which is capable of expressing CEA, and displays malignant potential and deserves great attention.

Objective To investigate the effects of gabapentin on high-voltage-activated calcium currents in dorsal root ganglion (DRG) neurons in mice with oxaliplatin-induced neuropathic pain (NP). Methods Pathogen-free male Kunming mice aged 6 weeks weighing 20-25 g were used in this study. NP was induced by injection of intraperitoneal oxaliplatin 3 mg/kg. Successful induction of NP was defined as the mechanical paw withdrawal threshold (MWT) measured at 3 d after oxaliplatin administration decreased to 40% of the baseline ( before administration of oxaliplatin). Forty-one mice in which NP was successfully induced were randomly divided into 2 groups: NP group ( n = 20) and gabapentin group (group G, n = 21 ). Another 10 normal mice served as control group (group C). At 3 days after oxaliplatin administration, gabapentin 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days in group G, while C and NP groups received the equal volume of normal saline.MWT to von Fray filament stimulation was measured immediately before and 1-3 days after gabapentin administration (T1-4). After the last measurement of MWT, bilateral L4.5 DRG was collected and neurons were isolated. The high-voltage-activated calcium currents were recorded using whole-cell patch-clamp technique. The peak current density and the voltage where half of the current was activated ( Va1/2 ) or inactivated ( Vi 1/2 ) were calculated. Results Compared with group C, MWT at T1-4 was decreased, the peak current density and Vi1/2 were significantly increased in group NP, and MWT at T1 was decreased in group G ( P ＜ 0.05). There was no significant difference in the peak current density, Vi1/2 and Va1/2 between C and G groups ( P ＞ 0.05). MWT at T2-4 was significantly increased, while the peak current density and Vi1/2 were significantly decreased in group G compared with group NP (P ＜ 0.05). Conclusion Gabapentin can reduce oxaliplatin-induced NP in mice through inhibiting high-voltage-activated calcium currents and promoting the inactivation of the channels in DRG neurons.

Objective To investigate the expressions of regulatory T cell (Treg) and interleukin-35 (IL-35) in patients with cholangiocarcinoma and to explore their clinical significance.Methods Flow-cytometry,PCR,Enzyme-linked immunosorbent assay (Elisa) and immunohistochemistry were used to detect the levels of Treg and IL-35 in peripheral blood and cholangiocarcinoma tissues in 42 patients with cholangiocarcinoma.Healthy volunteers were used as a control group.Result The percentage of Treg cells to CD4 + T cells in patients with cholangiocarcinoma was (5.6 ± 1.7) ％,while that in the normal control group was (2.9 ± 0.8) ％.There was a significant difference between the two groups (P ＜ 0.05).The plasma levels of IL-35 in patients with cholangiocarcinoma was (198.4 ± 81.4) pg/ml,while that in the normal control group was (33.7 ± 18.0) pg/ml.Again,a significant difference was observed between the two groups (P ＜ 0.05).In peripheral blood mononuclear cell,the IL-35 mRNA level was positively correlated with the plasma IL-35 level (p35,R =0.795,P ＜0.05;EBI3,R =0.812,P ＜ 0.05).Immunohistochemical studies showed that FOXP3 + tumor cells and Treg cells increased significantly in tumor tissues.Conclusion Overexpressions of Treg and IL-35 in peripheral blood and tumor tissues of patients with cholangiocarcinoma suggested that they may play important roles in the development of cholangiocarcinoma.

Objective To establish the ischemic precondition ([PC) model of PC12 cell line in vitro, and to explore the effect of nitric oxide (NO) on the IPC cerebral protection. Method PC12 cells were cultured and used for producing the model of ischemie precondition by the way of oxygen-glucose deprivation. Twenty dishes of cells were randomly divided into four groups (5 dishes for each group): control group, ischemic precondition group (IPC),non-ischemic precondition group (NIPC) and L-NAME treatment group (L-NAME). In control group, the cells were in-cubated with low glucose (<1 g/L) and2% FBS medium in normal oxygen; in IPC group, the cells were administrated with oxygen-glucose deprivation (OGD) for 6 hours, and then subjected with reperfuaion before OGD 15 hours; in NIPC group, the cells were treated the same as control group for 6 hours, and then subjected with reperfusion before OGD 15 hours; in L-NAME group, the cells received L-NAME (1 mmol/L) and cocultured for 30 minutes before OGD 6 hours, and then received the same treatment as the IPC group. To test whether the model was established, metabolic rate of MIT, LDH release were measured and the apoptosis rate was detected by flow cytometry following oxygen-glucose deprivation 15 hours. The activity of nitric oxide synthases (NOS) was as-sessed by biochemical assay. One-way ANOVA and LSD multiple comparison test were used to analyze differences among different groups, and P<0.05 was considered different. Results Compared with NIPC group, the metabolic rate of MTT increased (94.9%±35.1%, P<0.05), while LDH release and the cell apoptotic rate decreased significantly in IPC group (279.1%±28.1%, P<0.03). Compared with control group(100.0%± 13.5%),the activities of NOS increased both in NIPC and IPC groups (390.0%±14.6%, P<0.01;126.3% ±10.6%, P<0.01). Moreover, the apoptosis rates in each group (control group, IPC group, NIPC group and L-NAME group) were 5.90, 8.73, 38.62 and 11.73%,respectively. Conclusions IPC reduces the death and apoptosis rate of PC12 cell after oxygen-glucose deprivation injury. NO might be involved, but it is not the only factor.