Objective:To evaluate the effects of intranasal administration of glial cell line-derived neurotrophic factor (GDNF) on postoperative cognitive dysfunction in aged rats.Methods:Forty healthy Sprague-Dawley rats of both sexes, aged 21-23 months, weighing 480-600 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), operation group (group O), intranasal administration of low-dose GDNF group (group G1) and intranasal administration of high-dose GDNF group (group G2). Rats underwent exploratory laparotomy under anesthesia with chloral hydrate in O, G1 and G2 groups, while the rats in group S only received sham operation.The rats in group G1 and group G2 were intranasally treated with GDNF 25 and 50 μg (in 25 μl of PBS), respectively, and PBS 25 μl was nasally administered in group S and group O every day for 3 consecutive days after operation or sham operation.Morris water maze test was performed on days 3-7 after surgery, and then the rats were sacrificed, and hippocampal tissues were removed for determination of the expression of GDNF, high mobility group box 1 (HMGB1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), activated caspase-3 and Bax (by Western blot). Results:Compared with group S, the escape latency was significantly prolonged on days 5-7 after operation, the number of crossing the platform was reduced, time spent in the target quadrant was shortened, expression of GDNF was down-regulated, and expression of IL-1β, TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was up-regulated in group O, and the number of crossing the platform was reduced, time spent in the target quadrant was shortened, and expression of IL-1β and TNF-α was up-regulated in G1 and G2 groups ( P<0.05). Compared with group O, the escape latency was significantly shortened on days 5-7 after operation, the number of crossing the platform was increased, time spent in the target quadrant was prolonged, expression of GDNF was up-regulated, expression of TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was down-regulated in G1 and G2 groups, and IL-1β in hippocampi was down-regulated in group G1 ( P<0.05). Compared with group G1, the expression of TNF-α in hippocampi was down-regulated ( P<0.05), and no significant change was found in the other parameters mentioned above in group G2 ( P>0.05). Conclusions:Intranasal administration of GDNF can improve postoperative cognitive dysfunction, and the mechanism may be related to inhibiting neuroinflammatory responses and neuroapoptosis in aged rats.

Objective:To evaluate the effect of glucagon-like peptide 1 and glucose-dependent insulinotropic polypeptide receptor agonist DA-JC4 on postoperative neuroinflammatory responses in aged rats.Methods:Forty-five Sprague-Dawley rats, aged 21-23 months, weighing 530-630 g, provided by the Animal Experiment Center of Medical School of Zhengzhou University, were assigned into 3 groups ( n=15 each) using a random number table method: sham operation group (group S), operation group (group O) and DA-JC4 group (group G). Rats underwent exploratory laparotomy under anesthesia with chloral hydrate in O and G groups.In group G, DA-JC4 10 nmol/kg (dissolved in 1 ml of sterile normal saline) was intraperitoneally injected immediately after the end of operation and at 24 and 48 h after operation.Western blot was used to determine the expression of hippocampal Bax, Bcl-2, activated caspase-3, Beclin-1, microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ), high-mobility group box 1 protein (HMGB1), interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) on day 3 after surgery.The Morris water maze test was performed on days 14-18 after operation to assess the cognitive function. Results:Compared with group S, the escape latency was significantly prolonged on days 15-18 after operation in group O and on day 18 after operation in group G, and the number of crossing the original platform was reduced, the time spent in the target quadrant was shortened, the expression of activated caspase-3, Bax, LC3Ⅱ, HMGB1, IL-1β and TNF-α in hippocampi was up-regulated, and the expression of Bcl-2, LC3Ⅱ and Beclin-1 was down-regulated in O and G groups ( P<0.05). Compared with group O, the escape latency was significantly shortened, and the number of crossing the original platform was increased, the time spent in the target quadrant was prolonged, the expression of activated caspase-3, Bax, LC3Ⅱ, HMGB1, IL-1β and TNF-α in hippocampi was down-regulated, and the expression of Bcl-2, LC3Ⅱ and Beclin-1 was up-regulated in group G ( P<0.05). Conclusion:The mechanism by which DA-JC4 reduces postoperative cognitive dysfunction may be related to inhibiting neuroinflammatory responses in aged rats.

Objective To evaluate the effect of exercise training on heat shock protein 70 (HSP70) expression during endotoxin-induced acute lung injury (ALI) in rats.Methods Thirty-two SPF healthy male Sprague-Dawley rats,aged 8 weeks,weighing 175-220 g,were divided into 4 groups (n=8 each) using a random number table method:control group (group C),group ALI,low-intensity exercise training group (group ET1) and high-intensity exercise training group (group ET2).The rats in ET1 and ET2 groups received 2-and 4-week treadmill exercise training before establishing the ALI model,while the rats in C and ALI groups received no training.ALI was induced by intravenously injecting 5 mg/kg lipopolysaccharide via the tail vein in ALI,ET1 and ET2 groups,and the equal volume of normal saline was given instead in group C.The animals were sacrificed,and the lungs were harvested for microscopic examination of the pathological changes of lung tissues which were also scored and for determination of wet to dry weight ratio (W/D ratio),concentrations of total protein,interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) in bronchoalveolar lavage fluid (BALF),and expression of HSP70 and nuclear factor kappa B (NF-κB) in lung tissues by Western blot.Results Compared with group C,the W/D ratio and pathological changes of lung tissues were significantly increased,the concentrations of total protein,IL-1 β and TNF-α in BALF were increased,the expression of NF-κB was up-regulated (P＜0.05),and no significant change was found in HSP70 expression in group ALI(P＞0.05).Compared with group ALI,the W/D ratio and pathological changes of lung tissues were significantly decreased,the concentrations of total protein,IL-1β and TNF-α in BALF were decreased,the expression of HSP70 was up-regulated,and the expression of NF-κB was down-regulated in ET1 and ET2 groups (P＜0.05).Conclusion Exercise training can attenuate the endotoxin-induced ALI through relieving the inflammatory responses,which may be related to up-regulating HSP70 expression in the lung of rats.

Objective To evaluate the role of nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in remote ischemic preconditioning-induced reduction of lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.Methods Sixty-eight healthy male C57BL/6 mice,aged 6-8 weeks,weighing 22-26 g,were divided into 4 groups (n =17 each) using a random number table:control group (group C),ALI group,remote ischemic preconditioning group (group RIPC) and brusatol plus remote ischemic preconditioning group (group B+RIPC).Normal saline 100 μl was intratracheally instilled in group C.ALI was induced by intratracheal instillation of LPS 5 mg/kg in group ALI.Mice in group RIPC were subjected to 6 cycles of 5-min ischemia followed by 5-min reperfusion in the right hindlimbs using a tourniquet,and 1 h later the model of ALI was established.Nrf2 inhibitor brusatol 2 mg/kg (in 100 μl of 1％ dimethyl sulfoxide) was intraperitoneally injected every other day for 10 days prior to establishment of the ALI model in group B.Brusatol 2 mg/kg was intraperitoneally injected every other day for 10 days prior to establishment of the ALI model,and remote ischemic preconditioning was performed at 1 h before establishment of the ALI model in group B+RIPC.Seven mice in each group were selected at 24 h after establishment of the ALI model,and bronchoalveolar lavage fluid (BALF) was collected for determination of protein concentrations and neutrophil count.Mice were then sacrificed and lungs were removed for determination of lung water content,myeloperoxidase (MPO) activity,contents of interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α),and expression of Nrf2,HO-1 and high-mobility group box 1 protein (HMGB1) in lung tissues (by Western blot) and for examination of pathological changes (with a light microscope).Results Compared with group C,the lung water content,MPO activity,contents of IL-1β and TNF-α,and neutrophil count and protein concentrations in BALF were significantly increased,and the expression of Nrf2,HO-1 and HMGB1 was up-regulated in group ALI (P＜ 0.05).Compared with group ALI,the lung water content,MPO activity,contents of IL-1β and TNF-α,and neutrophil count and protein concentrations in BALF were significantly decreased,the expression of Nrf2 and HO-1 was up-regulated,and the expression of HMGB1 was down-regulated (P＜0.05),and the pathological changes were significantly attenuated in group RIPC.Compared with group RIPC,the lung water content,MPO activity,contents of IL-1β and TNF-α,and neutrophil count and protein concentrations in BALF were significantly increased,the expression of Nrf2 and HO-1 was down-regulated,and the expression of HMGB1 was up-regulated (P＜0.05),and the pathological changes were aggravated in group B+RIPC.Conclusion The activation of Nrf2/HO-1 signaling pathway is involved in remote ischemic preconditioning-induced reduction of LPS-induced ALI in mice.

Objective To investigate the role of transient receptor potential melastatin 7(TRPM7) in sevoflurane preconditioning for inhibiting hippocampal neurons apoptosis and inflammation response induced by oxygen-glucose deprivation(OGD).Methods Fifty SD rats of postnatal 1 d were selected for extracting hippocampal neurons and randomly divided into 5 groups,including the control group(C),sevoflurane preconditioning group (Sev),OGD group,Sev preconditioningt OGD group (Sev + OGD) and Sev preconditioning+ bradykinin+OGD group(combined group).After 1.5 h oxygen-glucose deprivation,reintroduction was performed,and then the normal culture was performed again for preparing the OGD model.Hippocampal neurons in the control group were normally cultured only;which in the Sev group conducted 2 ％ Sev preconditioning for 1 h;which in the OGD group only prepared the OGD model;which in the SEv+OGD conducted 2％ Sev preconditioning for 1 h,and prepared the OGD model after 24 h;which in the combined group was simultaneously added with bradykinin(final concentration 200μmol/L) in Sev preconditioning,other treatment was same to that in the Sev+OGD group.After 24 h normal culture,the mRNA and protein levels of TRPM7,apoptosis rate,survival rate,mRNA and supernatant protein levels of IL-1β and TNF-α of the hippocampal neurons were detected.Results Compared with the control group,hippocampal neurons mRNA and protein levels of TRPM7,apoptosis rate,mRNA and supernatant protein levels of IL-1β and TNF-α in the OGD group were significantly increased(P＜0.05),whereas the survival rate was significantly decreased (P ＜ 0.05).Compared with the OGD group,hippocampal neurons mRNA and protein levels of TRPM7,apoptosis rate,mRNA and supernatant protein levels of IL-1β and TNF-α in the Sev group were significantly decreased(P＜0.05),whereas the survival rate was significantly increased(P＜0.05).Compared with the Sev group,hippocampal neurons mRNA and protein levels of TRPM7,apoptosis rate,the mRNA and supernatant protein levels of IL-1β and TNF-α in the combined group were significantly increased(P＜0.05),whereas the survival rate was significantly decreased(P＜0.05).Conclusion Sev preconditioning can attenuate hippocampal neurons apoptosis and inflammatory response after OGD via alleviating the overexpression of TRPM7.

Objective To evaluate the relationship between α7 nicotinic acetylcholine receptor (α7nAChR) signaling pathway and regulatory T cells (Tregs) during vagus nerve stimulation-induced reduction of endotoxin-caused acute lung injury (ALI) in mice.Methods Clean-grade healthy male C57BL/6 mice,aged 6-8 weeks,weighing 22-25 g,were divided into 5 groups (n=10 each) using a random number table method:control group (group C),group ALI,vagus nerve stimulation group (group VNS),α-BGT group (group α-BGT) and vagus nerve stimulation plus α-BGT group (group VNS + α-BGT).After successful establishment of the model,the vagus nerve was stimulated for 30 s with a stimulus intensity of 0.5 mA,frequency of 20 Hz,an interval of 5 min,60 min in total.Sterile normal saline 100 μl was injected into the trachea,and the vagus nerve was only exposed but not stimulated in group C.In group ALI,the ALI model was established,and the vagus nerve was isolated but not stimulated.In group α-BGT,α-BGT 1 μg/kg was intraperitoneally injected,the ALI model was established 1 h later,and the vagus nerve was isolated but not stimulated.In group VNS+α-BGT,α-BGT 1 μg/kg was intraperitoneally injected,the ALI model was established 1 h later,and the vagus nerve was stimulated at 1 h after the end of establishment.Animals were sacrificed at 72 h after establishing the model,and lungs were removed for determination of lung water content,percentage of Tregs (using flow cytometry),myeloperoxidase (MPO) activity (by colorimetric assay),expression of α7nAChR (by Western blot) and contents of interleukin-10 (IL-10),transforming growth factor-β (TGF-β) and IL-1β (by enzyme-linked immunosorbent assay).Results Compared with group C,the lung water content,MPO activity and IL-1β content were significantly increased in the other four groups,the expression of α7nAChR was significantly down-regulated in ALI,α-BGT and VNS+α-BGT groups,the percentage of Tregs was significantly increased in group VNS,the IL-10 content was significantly decreased in ALI and α-BGT groups and increased in VNS and VNS+α-BGT groups,and TGF-β contents were significantly decreased in ALI,α-BGT and VNS+α-BGT groups and increased in group VNS (P＜0.05).Compared with group ALI,the lung water content,MPO activity and IL-1β content were significantly decreased,the expression of α7nAChR was up-regulated,and the percentage of Tregs and contents of IL-10 and TGF-β were increased in group VNS,and the TGF-β content in group α-BGT and contents of IL-10 and TGF-β in group VNS+α-BGT were significantly increased (P＜0.05).Compared with group VNS,the lung water content,MPO activity and IL-1β content were significantly increased,the expression of α7nAChR was down-regulated,and the percentage of Tregs and contents of IL-10 and TGF-β were decreased in α-BGT and VNS+α-BGT groups (P＜0.05).The contents of IL-1O and TGF-β were significantly higher in group VNS+α-BGT than in group α-BGT (P＜0.05).Conclusion Vagus nerve stimulation can activate α7nAChR signaling pathway and raise the percentage of Tregs,thus reducing ALI in mice.

Objective To investigate the effects of intrathecal dexmedetomidine administration on the development of morphine tolerance and spinal inflammatory responses.Methods Thirty-three male SD rats weighing 180～200 g were randomly divided into 3 groups (n=11):Saline group (group NS),Morphine group (group M) and Dexmedetomidine group (group Dex).Animals of group NS were intrathecally injected with 10 tμL of saline daily for seven days;Animals in group M were intrathecally injected with 15 μg of morphine daily for seven days;Animals in group Dex were intrathecally injected with a mixture of 15μg morphine and 1.5 μg dexmedetomidine daily for seven days.At 1,3,5 and 7 day of intrathecal injection,hot water tail-flick test were used to evaluate analgesic response to thermal stimuli.After the last episode of behavioral test,Western blot analysis was applied to determine the protein levels of Iba-1 (microglial marker),IL-1β,TNF-a and phospho-p38MAPK (p-p38) in the spinal cord.In addition,microglia in the spinal cord was immuno-stained with anti-Iba-1 antibody and the densities of microglia were calculated.Results In group M and Dex,the values of maximal possible effect (MPE) in tail-flick test decreased gradually along with repeated morphine administration (P＜0.05).Compared with group NS,the values of MPE in tail-flick test at 1,3,5 and 7 day of morphine tolerance were higher in group M (P＜0.05).Compared with group M,the values of MPE in tail-flick test at 3,5 and 7 day of morphine tolerance were higher in group Dex (P＜0.05).Compared with group NS,the spinal protein levels of Iba-1,IL-1L TNF-α and p-p38 as well as the density of Iba-1 positive cells in group M were increased (P＜0.05).However,Compared with group M,the of Iba-1,IL-1β,TNF-α and p-p38 as well as the density of Iba-1 positive cells were decreased(P＜0.05).Conclusion Intrathecal dexmedetomidine administration can attenuate morphine tolerance by inhibiting microglia-mediated inflammatory responses in the spinal cord.

Anterior cruciate ligament is the most important ligament to maintain the anterior and rotation stability of knee joint. Rupture of anterior cruciate ligament is one of the most common injuries of knee joint, and thus leads to knee instability and traumatic osteoarthritis. Anterior cruciate ligament reconstruction is usually performed to restore the anterior stability of knee joint, and is considered to reduce the secondary injury of medial meniscus, lateral meniscus and cartilage. Thus anterior cruciate ligament reconstruction can improve the function of knee joint. Traditional single bundle technique to reconstruct anterior cruciate ligament has been performed for many years. This technique can restore the anterior stability of knee joint and has excellent clini?cal results. Nearly 61%patients showed obvious radiographic osteoarthritis 20 years after anterior cruciate ligament reconstruction using bone-patella-bone graft. But, there is no agreement regarding to reduce the development of osteoarthritis after reconstruc?tion of anterior cruciate ligament. However, it has been reported that osteoarthritis would develop after reconstruction of anterior cruciate ligament in long term follow up study. Recently, with the further understanding of anatomy of biomechanics of anterior cru?ciate ligament, new techniques for anterior cruciate ligament reconstruction are developed, such as double bundle reconstruction, anatomic reconstruction and individual reconstruction. It remains controversial that whether these new technique can prevent the development of osteoarthritis after rupture of anterior cruciate ligament. Currently, no reconstruction technique for anterior cruci?ate ligament is perfect, and every technique has advantages and disadvantages. In terms of reducing the prevalence of osteoarthri?tis after reconstruction of anterior cruciate ligament, which technique is the best still remains unclear. New treatment and evalua?tion methods should be developed. In the future, not only the restoration of stability of knee joint should be considered, but also the articular cartilage contact kinematics including tibiofemoral joint and patellofemoral joint after anterior cruciate ligament recon?struction. Reduction of the development of osteoarthritis is an important topic after reconstruction of anterior cruciate ligament.

Objective To assess the morphological characteristics of bone marrow in patients of severe fever with thrombocytopenia syndrome ( SFTS) and its value in diagnosis.Methods The bone marrow morphology was retrospectively reviewed in 28 laboratory confirmed patients with SFTS from Zhoushan Hospital during January 2012 and December 2015.The correlation between bone marrow -derived macrophage and peripheral blood cells was analyzed with t test.Results All patients presented leukocytopenia and thrombocytopenia.Poor bone marrow hematopoietic function was observed in 23 patients (82%) showing granulocyte, erythrocyte and megakaryocyte hypoplasia , but no pathological hematopoietic disorder was observed.Eighteen patients (64%) had various degrees of increased amount of macrophage in the bone marrow; peripheral white blood cell count and platelets in patients with macrophage ≥0.5% were lower than those with macrophage <0.5%, and the difference was of statistical significance (t =3.836 and 4.499, P<0.01).Conclusion SFTS patients have characteristic bone marrow morphology , and bone marrow examination is beneficial for differentiation of SFTS from blood lymphatic system diseases and other virus infection.

Objective To analyze the relationship between cytotoxic T cells Perforin level, IFN-γand IL-10 in patients with chronic hepatitis B. Methods After a short-term cultivation of peripheral blood collected from 50 patients with chronic hepatitis B, a flow cytometry was employed to detect the levels of Per-forin, IFN-γ and IL-10 in CD8~+ cells to compare with those of the health donors the HBV DNA copies in their blood were also measured by RT-PCR to analyze the relationship with Perforin, IFN-γ, and IL-10 in CD8~+ cells. Results In patients with chronic hepatitis B, the levels of Perforin, IFN-γ and IL-10 in CD8~+ ceils were (5.30 ± 2.62)%, (4.05 ± 2.25) % and (0.77 ± 0.50) %, respectively, which were statistical-ly lower than health donors(the t values were 4.50, 4.56 and 4.20 respectively; P < 0.01) ; Of 26 cases of chronic hepatitis B patients with HBeAg-positive Pefforin and IFN-γ in CD8~+ T cells were (4.54 ± 1.93) % and (3.32 ± 1.59)%, respectively, significantly lower than those of the 24 chronic hepatitis B patients with HBeAg-negative (the t values were 2.22 and 2.54, respectively; P <0.05) ; And the levels of Perforin, IFN-γ had a negative relation with the HBV DNA copies (the coefficient correlations-0. 539 and-0. 340; P < 0.01 and P < 0.05). Conclusion In patients with chronic hepatitis B, the reduced levels of Pefforin, IFN-γ and IL-10 may be related with the long-term existence of HBV, and the protracted course of disease.