Objective To evaluate the effect of remote limb ischemic postconditioning (RLIP) combined with electro-acupuncture postconditioning (EAP) on focal cerebral ischemia-reperfusion (I/R) injury in rats.Methods Seventy male Sprague-lDawley rats,weighing 220-250 g,were randomly assigned into 5 groups (n =14 each) using a random number table:sham operation group (group S),group I/R,RLIP group,EAP group and RLIP combined with EAP group (group RLIP +EAP).Focal cerebral I/R was induced by middle cerebral artery occlusion (MCAO).MCAO was maintained for 120 min.The animals were subjected to 3 cycles of 15 min ischemia of bilateral hind limbs followed by 30 s reperfusion starting from onset of reperfusion.Dazhui and Baihui acupoints were stimulated with electric stimulator (frequency 15 Hz/2 Hz,intensity 1 mA) for 30 min starting from onset of reperfusion.Neurological deficit scores (NDSs) were evaluated at 24 h of reperfusion.The animals were then sacrificed and brains were removed to measure the infarct size and for microscopic examination of pathological changes in hippocampal CA1 region.The number of survival neurons was counted.The apoptosis in neurons was determined by TUNEL.Apoptosis index was calculated.Results Compared with group S,the NDSs,percentage of cerebral infarct size and AI were significantly increased,and the number of survival neurons was decreased in the other four groups.Compared with group I/R,the NDSs,percentage of cerebral infarct size and AI were significantly decreased,and the number of survival neurons was increased in EAP,RLIP and RLIP+EAP groups.Compared with RLIP and EAP groups,the NDSs,percentage of cerebral infarct size and AI were significantly decreased,and the number of survival neurons was increased in group RLIP+ EAP.Conclusion The combination of RLIP and EAP provides better efficacy than either alone in attenuating focal cerebral I/R injury in rats.

AIM: To investigate the effect of sulfated cholecystokinin octapeptide (CCK -8 ) on TNF -α induced IL - 6 mRNA expression, NF - κB activation in the rat fibroblast - like synovial cell strain RSC - 364 and its possible receptor mechanisms. METHODS: RSC -364 cells were stimulated with TNF - α( 10 μg/L) in the presence or absence of sCCK- 8( 10-8 - 10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L). IL -6 and CCK receptor A/B (CCK- AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction (RT- PCR) at 3 h after stimulation, and nuclear factor - κB (NF - κB) binding activity was analyzed by electrophoretic mobility shift assay (EMSA) at lh after stimulation. At 30 min of stimulation the IκB protein level in cytoplasma was measured by Western blotting. RESULTS: Both CCK - AR and CCK - BR were constitutively expressed on RSC - 364. sCCK - 8, at concentrations from 10-8 mol/L to 10 -6 mol/L, significantly increased IL - 6 mRNA expression, CCK - AR and CCK - BR mRNA expression, NF - κB binding activity and IκB protein degradation. The effects of sCCK - 8 on NF - κB activity and IκB degradation level were attenuated by CCK receptor antagonist proglumide. CONCLUSION: sCCK - 8 upregulats TNF - α- induced IL - 6 mRNA expression by NF - κB pathway through its receptor on rat synoviocytes, suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.

Objective To explore the influence of high fat diet on learning and memory,as well as the alteration of the number of neurons and morphology of dendritic spines in rat hippocampi.Methods 24 male adult SD rats were randomly assigned to high fat diet group or control group.The rats were fed with high-fat diet or standaM laboratory rodent chow diet for 12 weeks.Learning and memory were tested by Morris water maze and object recognition tests, and mood and motor ability were tested by open field tests.Golgi staining detected dendritic spine density of hippocampal neurons, and Nissl staining was used to observe the number of hippocampal neurons and pathological changes.Results High-fat diet induced rat spatial learning deficits, which was demonstrated by the prolonged escape latency ((38.50±9.70) s, (20.08±7.35) s, (19.96± 10.56) s, (22.75± 12.51) s, (14.56±4.82) s) compared with the control ((33.61±12.41) s, (14.25±7.89) s, (15.06±7.59) s, (5.53±2.81) s, (4.7± 1.58) s).The spatial memory deficits demonstrated that the latency reaching platform ((30.46± 21.43) s) was prolonged compared with control ((5.18± 1.33)s).The working memory was impaired, which was demonstrated by the prolonged escape latency compared with control group (P＜ 0.05).Discrimination index lowered than control group ((0.67±0.12) vs (0.81±0.08)), and the difference was significant (P=0.038), but no anxiety behaviors were observed(P=0.461).The neuron number of hippocampal neurons and dendritic spine density were significantly lowered than those in the control group((209.73±24.29) vs (262.2±18.94), (17.9±2.84) vs (21.93±2.56) ,respectively) (P＜0.05).Conclusion Intake of high-fat diet can impair learning and memory in rats, as well as decrease the number of neurons and the density of dendritic spines in the hippocampus.

Objective To explore the mechanism and safety of miltidrug-resistance gene 1(MDR1)transfection into the bone marrow mononuclear cells of rabbit in vitro with the adenovirus vector promoted by ultrasonic microbubble.MethodsBone marrow mononuclear cells of rabbits were collected and divided into 5 groups after cultured in the 6-well plate according to the different experimental conditions(MDR1 gene was transferred into the cells with or without ultrasound irradiation and microbubbles)：conventional culture group (A)，Ad5-MDR1 group(B)，Ad5-MDR1+ultrasound irradiation group(C)，Ad5-MDR1+microbubbles group(D)，Ad5-MDR1+ultrasound irradiation+microbubbles group(E).The positive transfection rate of MDR1 gene in mononuclear cells of different groups were tested by flow cytometry,and the survival rate of cells in different periods were tested by trypan blue exclusion method.Moreover,the appearance and ultramicrostructure of cells were observed by electronmicroscope.Results①The transfection rate of MDR1 gene in different groups were 0.39%±0.11%，5.03%±0.35%，4.93%±0.38%，5.25%±0.80%and 19.93%±1.51%respectively.The transfection rate of MDR1 gene in group E was higher than those in other groups(P＜0.05).②Compared with those in control groups(group A，B，C and D)，the transfection rate in group E was significant raised by ultrasound irradiation and microbubbles.However,there were no significant difference in survival rate of cells between the five groups(P＞0.05).③After ultrasonic irradiation，there were transient holes on the cell membrane，which could disappear after irradiation by ultrasound for 24 hours.And the temporary swelling of organelles was reversible.Conclusions Microbubbles irradiated by ultrasound can cause small transient holes on eell membrane and increase permeability of it，and enhance the transfection of MDR1 gene in bone marrow mononuclear cells with the adenovirus vector，which is safe and available.

Objective To observe the effect of autophagy on paclitaxel-induced CaSki cell death through the regulation of the expression of autophagy gene Beclin1, and to explore the interaction and relationship between autophagy and apoptosis. Methods Eukaryotic expression vector pcDNA3.1-Beclin1 and RNA interference vector pSUPER-Beclin1 were transfected into human cervical cancer CaSki cells in vitro and screened for stable expression cell lines. The formation of autophagic vacuoles was observed with an electronic microscope. The expression of Beclin1 and LC3 was measured by Western blot. After being treated with paclitaxel, the change of cell proliferation was assessed by MTT assay, the percentage of apoptotic cells and autophagic cells were analyzed by flow cytometry. Results A lot of autophagic vacuoles were observed in pcDNA3.1-Beclin1 cells by electronic microscopy. Beclin1 and LC3 protein expression was up-regulated in CaSki cells transfected with pcDNA3.1-Beclin1, and was inhibited in cells transfected with pSUPER-Beclin1. MTT assay revealed the survival rate of CaSki cells was significantly decreased after being transfected with pcDNA3.1-Beclin1. After being treated with paclitaxel, the percentages of apoptotic cells and autophagic cells were both increased in pcDNA3.1-Beclin1 group compared with that of the blank control group especially the increase of apoptosis was particularly evident. Conclusion Autophagy and apoptosis have different roles in the process of paclitaxel-induced cervical cancer CaSki cell line death. Overexpression of Beclin1 in CaSki cells may enhance the apoptosis induced by paclitaxel.

Objective To investigate the status of nutritional risk and nutritional support in general surgery patients, and to explore their association with postoperative complications and length of hospital stay.Methods From January 2014 to February 2015, 853 inpatients in general surgical wards in the Second Hospital of Hebei Medical University were enrolled.Nutritional Risk Screening 2002 (NRS 2002) was used to estimate nutritional status of patients.The patients were divided into 2 groups based on whether they received nutritional support.The length of hospital stay in days and postoperative complications were recorded.The association of nutritional risk and nutritional support with complications and length of hospital stay were analyzed.Results In the 853 surgery patients, the prevalence of nutritional risk was 31.1％ (265/853) and that of malnutrition was 5.4％ (46/853).The incidence of postoperative complications was 14.2％ (121/853).The patients with nutritional risk had a significantly higher incidence of postoperative complications compared to those without nutritional risk [29.8％ (79/265) vs.7.1％ (42/588) , P ＜ 0.000] , and a longer hospital stay [(12.5 ±6.4) days vs.(4.2 ±3.9) days, P ＜0.001].In the 853 patients, 27.3％ (233/853) received nutrition support.In the patients with nutritional risk, those on nutritional support had a significantly lower incidence of complications compared with those not on nutritional support [16.7％ (32/192) vs.64.4％ (47/73), P＜0.05] and shorter hospital stay [(7.5±4.6) days vs.(16.3±8.5)days, P ＜ 0.05].Conclusions According to NRS 2002 result, a fairly high percentage of general surgery patients may have nutritional risk.Patients with decreased body mass, less dietary intake, and at higher age may be more likely to have nutritional risk.Nutritional risk may be associated with a higher incidence of postoperative complications and longer hospital stay.Patients at nutritional risk appear to be more likely to benefit from nutritional support.

Objective To analyze the cognition and demand of the residency training program among clinical medical students to provide scientific basis for the promotion of this policy.Methods From May 24 to 25,2015,a self-designed questionnaire survey was conducted on 645 undergraduate clinical medical students,who came from Zhengzhou University,Xinxiang Medicine College,Henan University.Data of cognition and demand of the residency standardized training were statistically analyzed using SPSS 22.0 software.Enumeration data were analyzed by chi-square test and abnormal distribution data based on Wilcoxon rank sum test.Results 616 valid questionnaires were recovered.259 undergraduate clinical medical students (42.00％) did not quite understand the resident standardized training policy and 225 (36.50％) did not known the policy at all.Before and after clinical practice,the cognition of the residency standardized training had statistically significant difference (x2=87.596,P=0.000),the knowledge of the policy (x2=10.939,P=0.012) and the degree ascending alleviate employment pressure (x2=29.349,P=0.000) were the main influencing factors of the clinical medical students' choices after graduation.Conclusion Improving medical students' understanding of the significance of medical education after graduation,and the integration of professional degree training and residency standardization training effectively helps to promote implementation of the policy.

AIM:To investigate the effect of sulfated cholecystokinin octapeptide(CCK-8) on TNF-? induced IL-6 mRNA expression,NF-?B activation in the rat fibroblast-like synovial cell strain RSC-364 and its possible receptor mechanisms.METHODS:RSC-364 cells were stimulated with TNF-?(10 ?g/L) in the presence or absence of sCCK-8(10-8-10-6 mol/L) or/and CCK receptor antagonist proglumide(2 mg/L).IL-6 and CCK receptor A/B(CCK-AR/CCK/BR) mRNA expression were assayed by reverse transcription polymerase chain reaction(RT-PCR) at 3 h after stimulation,and nuclear factor-?B(NF-?B) binding activity was analyzed by electrophoretic mobility shift assay(EMSA) at 1h after stimulation.At 30 min of stimulation the I?B protein level in cytoplasma was measured by Western blotting.RESULTS:Both CCK-AR and CCK-BR were constitutively expressed on RSC-364.sCCK-8,at concentrations from 10-8 mol/L to 10-6 mol/L,significantly increased IL-6 mRNA expression,CCK-AR and CCK-BR mRNA expression,NF-?B binding activity and I?B protein degradation.The effects of sCCK-8 on NF-?B activity and I?B degradation level were attenuated by CCK receptor antagonist proglumide.CONCLUSION:sCCK-8 upregulats TNF-?-induced IL-6 mRNA expression by NF-?B pathway through its receptor on rat synoviocytes,suggesting its possible regulatory role in the pathogenesis of rheumatoid arthritis.

AIM: To investigate the inhibitory effect s of a synthetic CRE-transcription factor decoy oligodeoxynucleotide (CRE-decoy ODN) on the upregulation of the expression of cholecystokinin (CCK) and fosB mRN A induced by chronic morphine administration in SK-N-SH cells. METHODS: The CRE cis-element, TGACGTCA, was palindromic, a sy nthetic single-stranded phosphorothioate oligodeoxynucleotide composed of the CR E sequence self-hybridizes to form a duplex/hairpin. The CRE-palindromic decoy a nd control oligodeoxynucleotides were added to the medium (1 h before exposure t o morphine) at 150 nmol/L in the presence of cationic lipid DOTAP. After the cel ls were treated with 100 ?mol/L morphine for 48 h, 10 ?mol/L naloxone was use d for 15 min. The effects of CRE-decoy ODN on the DNA-binding activity of CREB, the expression of CCK and fosB mRNA were detected by electrophoresis mobi lity shift assay (EMSA) and RT-PCR, respectively. The stability of cell-incorpo rated [ 32P]-labeled CRE-decoy ODN was extracted with phenol:chloroform a nd then subjected to 20% nondenaturing polyacrylamide gel electrophoresis and au toradiography. RESULTS: Chronic morphine administration and acute naloxone-prec ipitated withdrawal significantly activated the DNA-binding activity of CREB and the expression of CCK and fosB mRNA in SK-N-SH cells. The CRE-decoy ODN pen etrated into the cells, specifically downregulated these indexes. CONCLUSIONS: CRE-decoy ODN can significantly downregulates the e xpre ssion of CCK and fosB mRNA through specifically suppressing the DNA-binding activity of CREB activated by chronic morphine administration in SK-N-SH cells.

Objective To evaluate the rat model of type 2 diabetes mellitus ( T2DM) associated with depres-sion by body weight, fasting blood glucose, manifestations, and open field test.Methods The T2DM rat model was induced by high fat diet and low dose of STZ injection, and in addition, the T2DM rats were made restraint stress for 21 days.32 Wistar rats were randomly divided into three groups: normal group ( group N) , T2DM group ( group T)and T2 DM with depression group ( group T +D) , with 8 rats in each group.After the model was established, to measure the body weight, fasting blood glucose at day 0, 7, 14, and 21, and observe the gross manifestations, drink-ing and diet, feces, urine, and mental state, and test the rat depression by open field test.Results After establish-ment of the T+D rat model, the rats in group T+D showed some symptoms, including messy dark and gloomy hair, slow movement, increasing drinking, diet, feces and urine and mental fatigue.At day 0, 7, 14, and 21, compared with the group N, the body weight of the group T and group T+D were decreased, showing a significant difference ( P<0.05;P<0.01).At day 0, 7, 14, and 21, compared with the group N, and the fasting blood glucose in the groups T and T+D were increased, with a significant difference ( P<0.01 ) .After 21 days of restraint stress, the fasting blood glucose in the group T+D was significantly higher than that in the group T ( P<0.01 ) .Compared with the group N, the total movement distance in 5 minutes in the group T+D was reduced, but without a significant differ-ence (1532.6 ±126.8 cm vs.940.5 ±208.3 cm, P>0.05).Compared with the group N, the movement speed in 5 minutes in the group T was significantly slower than that in the group T , with a significant difference ( 5.1 ±0.4 cm/s vs.2.9 ±0.6 cm/s, P<0.05 ) , and even more slower than that in the group T +D, with a significant differ-ence (5.1 ±0.4 cm/s vs.2.4 ±0.5 cm/s, P<0.01) .Conclusions A rat model of type 2 diabetes mellitus as-sociated with depression has been successfully established by high fat diet and injection of low dose streptozotocin in combination with restraint stress for 21 days.This rat model is useful for further relevant studies.