Objective:To evaluate the role of glutathione S-transferase μ1 (GSTM1) expression in mild hypothermia-induced mitigation of cerebral ischemia-reperfusion (I/R) injury and the relationship with microglial polarization.Methods:Eighty clean-grade healthy male Sprague-Dawley rats, aged 8 weeks, weighing 260-280 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (S group), cerebral I/R group (I/R group), mild hypothermia group (H group), and GSTM1 inhibitor + mild hypothermia group (IH group). The rat model of cerebral I/R injury was prepared using the filament occlusion method. The filament was removed to restore blood flow after the left middle cerebral artery was blocked for 2 h, and the rats′ brain and rectal temperature were maintained at 36-37 ℃ during the period. The vessels were only isolated and ligated without occlusion in S group. In H group, the entire body was wiped with 75% ethanol immediately after removing the filament, and the brain and rectal temperatures were maintained at 32-33 ℃ for 3 h, and the other procedures were the same as those previously described in I/R group. In IH group, GSTM1 inhibitor itaconic acid 8.6 mg/kg was intraperitoneally injected at 24 and 1 h before developing the model, and the other procedures were the same as those previously described in H group. Neurological deficits were evaluated using a modified neurological severity score (mNSS) at 24 h of reperfusion, and then the animals were sacrificed and the brains were removed for observation of cerebral infarction (by TTC staining) and for determination of the expression of GSTM1, M1-type microglial marker inducible nitric oxide synthase (iNOS), and M2-type microglial marker arginase-1 (Arg-1) (by Western blot), expression of GSTM1, iNOS and Arg-1 mRNA (quantitative real-time polymerase chain reaction) and contents of interleukin-6 (IL-6), IL-10, tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) (by enzyme-linked immunosorbent assay). Results:Compared with S group, the mNSS and percentage of cerebral infarct size were significantly increased, and the expression of iNOS and Arg-1 protein and mRNA was up-regulated, the expression of GSTM1 and mRNA was down-regulated, and the contents of IL-6, TNF-α, IL-10 and TGF-β were increased in the other three groups ( P<0.05). Compared with I/R group and IH group, the mNSS and percentage of cerebral infarct size were significantly decreased, and the expression of iNOS protein and mRNA was down-regulated, the expression of Arg-1 protein and mRNA and GSTM1 was up-regulated, the contents of TNF-α and IL-6 were decreased, and the contents of TGF-β and IL-10 were increased in H group ( P<0.05). Conclusions:Up-regulated expression of GSTM1 is involved in mild hypothermia-induced mitigation of cerebral I/R injury, which is associated with inhibition of microglial polarization toward the M1 phenotype and promotion of polarization toward the M2 phenotype.

Objective:To study the infection of human rhinovirus, respiratory syncytial virus and human adenovirus in Jinan from April 2018 to March 2019, and analyze epidemiological characteristics of human adenovirus.Methods:All of 1969 nasopharyngeal swabs were collected from hospitalized patients with acute respiratory tract infections in The Fourth People’s Hospital of Jinan, Qilu Children′s Hospital of Shandong University, Peoples Hospital of Zhangqiu District from April 2018 to March 2019, fluorescence quantitative PCR was used to detect the positive rate of human rhinovirus, respiratory syncytial virus and human adenovirus. Seven adenovirus positive samples were isolated and examined by sequencing, then we determined adenovirus type, constructed gene phylogenetic trees for analysis.Results:Of the 1969 samples, 242 were positive, the total positive rate was 12.30% (242/1969), the positive rate was 3.00% (59/1969) for rhinovirus, 6.30% (124/1969) for respiratory syncytial virus (RSV), and 3.86% (76/1969) for adenovirus. There was no significant difference in the detective rate of rhinovirus in different age groups (Fisher′s exact test value =8.376, P=0.720), the positive rates of RSV and adenovirus in different age groups was statistically significant ( χ2=19.28, 12.16; P=0.001, 0.016). There was a statistically significant difference in the positive rate of adenovirus between different sexes ( χ2=14.33, P<0.001), and there was no statistically significant difference in the positive rate of rhinovirus and RSV between males and females ( χ2=0.30, 2.90, P=0.862, 0.089). Comparing the positive rates of viral nucleic acid in different months, we found that the positive rate of rhinovirus, RSV and adenovirus separately reached the highest in October, December and November (8.61%, 26.50% and 8.84%). We constructed a gene phylogenetic tree after seven positive samples of adenoviruses were sequenced, by the molecular typing method we detected that seven adenovirus-positive samples were all HAdV-2 type. Conclusions:By comparing the epidemiological trends of human rhinovirus, RSV and human adenovirus in Jinan from April 2018 to March 2019 in different ages, genders, and months, providing reference basis for the early prevention and clinical treatment of acute respiratory tract infection.

Objective:To evaluate the relationship between M2-type microglia-derived exosomes (M2-exo) and neuronal oxygen-glucose deprivation and restoration (OGD/R) injury in mice.Methods:Mouse neuroblastoma cells (N2a cells) and BV2 microglia were cultured in vitro, and BV2 microglia were activated to M2 type using 20 ng/ml IL-4, and M0-type microglia-derived exosomes (M0-exo) and M2-exo were extracted.N2a cells were divided into 4 groups ( n=23 each) using the random number table method: control+ M0-exo group (C+ M0 group), control+ M2-exo group (C+ M2 group), OGD/R+ M0-exo group (O+ M0 group) and OGD/R+ M2-exo group (O+ M2 group).M0-exo and M2-exo (final concentration 100 μg/ml) were added in C+ M0 and C+ M2 groups, respectively, and the cells were incubated for 24 h. M0-exo and M2-exo (final concentration 100 μg/ml) were added at 3 h after oxygen and glucose deprivation, and then the cells were incubated for 24 h in O+ M0 and O+ M2 groups, respectively.N2a cell viability was measured by the CCK-8 method, and the severity of cell damage was assessed using the lactic dehydrogenase (LDH) release rate.The expression of Bax and Bcl-2 protein and mRNA was detected by quantitative real-time polymerase chain reaction and Western blot. Results:Compared with C+ M0 group, no significant changes were found in N2a cell viability, LDH release rate, Bax/Bcl-2 ratio and Bax mRNA/Bcl-2 mRNA ratio in C+ M2 group ( P>0.05), and N2a cell viability was significantly decreased, and the LDH release rate, Bax/Bcl-2 ratio and Bax mRNA/Bcl-2 mRNA ratio were increased in O+ M0 group ( P<0.05).Compared with C+ M2 group, the N2a cell viability was significantly decreased, and the LDH release rate, Bax/Bcl-2 ratio and Bax mRNA/Bcl-2 mRNA ratio were increased in O+ M2 group ( P<0.05).Compared with O+ M0 group, N2a cell viability was significantly increased, and LDH release rate, Bax/Bcl-2 ratio, and Bax mRNA/Bcl-2 mRNA ratio were decreased in O+ M2 group ( P<0.05). Conclusions:M2-exo exerts an endogenous protective effect during OGD/R in mouse neurons, which may be related to the inhibition of cell apoptosis.

Objective:To evaluate the role of hsa_circ_0025853 in hypothermia-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury to neuroblastoma cells (SK-N-SH cells). Methods:SK-N-SH cells were cultured in vitro to the logarithmic phase and divided into 4 groups ( n=20 each) using a random number table method: control group (group C), group OGD/R, hypothermia group (group H) and hsa_circ_0025853 small interfering RNA (siRNA) plus hypothermia group (group S+ H). The cells were cultured in normal culture atmosphere in group C. The cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply for 24 h in group OGD/R.The cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply for 24 h at 32 ℃ in group H. The cells were transfected with siRNA at 48 h before establishing OGD/R model, and the other treatments were similar to those previously described in group H. At 24 h of restoration of oxygen-glucose, cell viability was measured by Cell Counting Kit-8, apoptosis rate were measured by flow cytometry.the expression of dynamin-related protein 1 (Drp1) mRNA and hsa_circ_0025853 was detected by quantitative real-time-polymerase chain reaction, and the expression of Drp1 protein was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of hsa_circ_0025853 was down-regulated, and the expression of Drp1 protein and mRNA was up-regulated in the other 3 groups ( P<0.05). Compared with group OGD/R, the cell viability was significantly increased, the apoptosis rate of cells was decreased, the expression of hsa_circ_0025853 was up-regulated, and the expression of Drp1 protein and mRNA was down-regulated in H and S+ H groups ( P<0.05). Compared with group H, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of hsa_circ_0025853 was down-regulated, and the expression of Drp1 protein and mRNA was up-regulated in group S+ H ( P<0.05). Conclusion:The mechanism by which hypothermia alleviates the OGD/R injury to SK-N-SH cells is related to the up-regulation of hsa_circ_0025853 expression, thus reducing the expression level of Drp1.

Objective:To evaluate the effect of electroacupuncture (EA) preconditioning on expression of cortical Ubc9 during cerebral ischemia-reperfusion (I/R) in rats.Methods:A total of 80 healthy clean-grade male Sprague-Dawley rats, aged 8-12 weeks, weighing 200-250 g, were divided into 4 groups ( n=20 each) using a random number table method: sham operation group (group S), group I/R, EA preconditioning group (group E) and sham EA group (group SE). Blood vessels were only exposed, without occlusion in group S. In the other three groups, the cerebral I/R model was established by middle cerebral artery occlusion using suture-occluded method, and the suture was removed after 2-h occlusion to restore the perfusion in anesthetized rats.EA was performed at 5 days before establishing the model in group E and group SE.Baihui acupoints were stimulated with an electric stimulator (2/12 Hz disperse-dense waves, intensity 1 mA) for 30 min once a day for 5 consecutive days, and the model was established at 24 h after the last stimulation.EA was performed at the points 1 cm lateral to the acupoints of Baihui, and the other operating parameters were the same as those previously described in group E. Neurological deficit scores (NDSs) were evaluated at 24 and 48 h of reperfusion.Then the rats were sacrificed, and tissues in the ischemic penumbra of cerebral cortex were obtained for determination of cell apoptosis (by TUNEL) and expression of Ubc9 and conjugated small ubiquitin-like modifier (SUMO) 2/3 (by Western blot). The apoptosis rate was calculated. Results:Compared with group S, NDSs at 24 and 48 h of reperfusion and apoptosis rate were significantly increased, and the expression of Ubc9 and conjugated SUMO2/3 was up-regulated in the other three groups ( P<0.05). Compared with group I/R and group SE, NDSs at 24 and 48 h of reperfusion and apoptosis rate were significantly decreased, and the expression of Ubc9 and conjugated SUMO2/3 was up-regulated in group E( P<0.05). Conclusion:The mechanism by which EA preconditioning reduces cerebral I/R injury is related to up-regulating Ubc9 expression and thus enhancing SUMO2/3ylation in rats.

Objective: To compare the efficacy of different decompressions treatment combined with posterior cervical fusion and internal fixation treatment on Chiari I malformation associated with syringomyelia and atlantoaxial instability. Methods: From January 2016 to October 2017, 36 cases of Chiari I malformation associated with syringomyelia and atlantoaxial instability in Linfen People's Hospital were selected in the research.The patients were divided into two groups according to random number table method, with 18 cases in each group.The observation group was given decompression of posterior fossa of small bone window + cerebellar tonsillectomy + posterior cervical fusion and internal fixation treatment.The control group was given traditional decompression of posterior fossa + posterior cervical fusion and internal fixation treatment.The efficacy, complications occurred within 1 year after operation, JOA score and spinal cord cavity size at different time points before and after operation of the two groups were compared. Results: The total effective rate of the observation group [94.44%(17/18)] was slightly higher than that of the control group [88.89%(16/18)] (χ²=1.957, P>0.05). The total effective rate of the observation group at 1 year after operation was 88.89%(16/18), which was significantly higher than that of the control group [72.22%(13/18)] (χ²=3.498, P<0.05). The incidence rate of complications of the observation group at 1 year after operation was 11.11%(2/18), which was significantly lower than that of the control group [38.89%(7/18)](χ²=11.685, P<0.05). The JOA scores at discharge[(15.97±1.25)point] and 1 year after operation[(15.53±1.19)point] of the observation group were significantly higher than those of the control group[(14.21±1.18)point, (14.06±1.15)point](t=3.815, 3.706, all P<0.05). The spinal cord cavity size at discharge [(2.13±0.64)mm] and 1 year after operation[(2.28±0.69)mm] of the observation group were significantly higher than those of the control group [(3.41±0.76)mm, (3.45±0.78)mm](t=5.743, 5.511, all P<0.05). Conclusion The long-term efficacy of decompression of posterior fossa of small bone window + cerebellar tonsillectomy + posterior cervical fusion and internal fixation treatment on Chiari I malformation associated with syringomyelia and atlantoaxial instability is remarkable, the recovery of spinal cord function and syringomyelia is well and the postoperative complications incidence is relatively low, which is worthy of popularization and application.

Objective To evaluate the role of α7 nicotinic acetylcholine receptor (α7nAChR) in postoperative cognitive dysfunction in aged rats with tibial fracture. Methods One hundred and fifty clean-grade healthy male Sprague-Dawley rats, aged 18-22 months, weighing 440-580 g, were divided into 5 groups ( n=30 each ) using a random number table method: control group ( group C ) , sham operation group ( group S) , tibial fracture group ( group T) , normal saline group ( group N) and α7nAChR agonist PUN282987 group (group P). Group C received no treatment. Ten percent chloral hydrate 0. 4 ml/100 g was injected intraperitoneally in group S. Group T underwent tibial fracture. PUN2829872. 4 mg/kg was in-traperitoneally injected at 5 min before tibial fracture in group P . The equal volume of normal saline was giv-en at 5 min before tibial fracture in group N. Morris water maze test was performed at day 7 after surgery. At days 1, 3 and 7 after surgery, the pathological changes of the hippocampal CA3 region were observed by haematoxylin and eosin staining, and the expression of α7nAChR, choline acetyltransferase ( ChAT ) , tumor necrosis factor-α( TNF-α) and interleukin-1β( IL-1β) in the hippocampal CA3 region was measured by Western blot. Results Compared with group C, the postoperative escape latency and swimming dis-tance were significantly prolonged, and the expression of α7nAChR, ChAT, TNF-α and IL-1β was up-regulated at each time point after operation in T, N and P groups ( P＜0. 05) , and no significant change was found in the parameters mentioned above in group S ( P＞0. 05) . Compared with group T, the postoper-ative escape latency and swimming distance were significantly shortened, and the expression of α7nAChR and ChAT was up-regulated and the expression of TNF-α and IL-1β was down-regulated at each time point after operation in group P ( P＜0. 05) , no significant change was found in the parameters mentioned above in group N ( P＞0. 05) , and the pathological changes of the hippocampal CA3 region were significantly at-tenuated in group P. Conclusion α7nAChR antagonism is involved in the development of postoperative cognitive dysfunction in aged rats with tibial fracture.

Objective To evaluate the effect of extremity ischemia preconditioning on lung protection in patients undergoing pulmonary lobectomy,and to further explore its possible mecha-nisms.Methods Forty patients (male 26 and female 14,BMI 20-28 kg/m2,ASA Ⅰ or Ⅱ,aged 45-64 years old)scheduled for elective pulmonary lobectomy via a thoracoscope,were randomly divided into two groups:extremity ischemic preconditioning group (group P)had the left leg roots tied with a tourniquet,lower extremity blood flow aerated blocked for 5 min,then deflated to restore blood flow for 5 min,the cycle was repeated three times;control group (group C)had the left lower extremity tied but not charged for 30 min.All the patients underwent intravenous-inhaled composite anesthesia. Arterial and venous blood samples were taken after admission to the operating room,at 6 h,12 h and 24 h after operation.The alveolar-arterial oxygen pressure difference (PA-aO2),respiratory index(RI) and oxygenation index (OI)were calculated.The level of TLR4 was measured.The pulmonary com-plications within 48 h after operation were recorded.The adverse reactions of the left lower extremity of the group P were recorded.Results Compared with the admission to the operating room,the ex-pression of TLR4 was significantly increased in the two groups at 6 h,12 h and 24 h after operation, but the expression of the group P was significantly lower than that of group C (P<0.05).Compared with the admission to the operating room,PA-aO2and RI were significantly increased,and OI was sig-nificantly decreased in the two groups at 6 h,12 h and 24 h after operation (P<0.05).Compared with group C,PA-aO2and RI were significantly decreased,and OI was significantly increased in group P (P<0.05).There was no statistically significant difference in the incidence of pneumonia and ate-lectasis in the two groups after operation,and no respiratory failure was observed in both groups.The left lower extremity of the group P had no adverse reactions,such as rupture of skin,thromboembo-lism,and nerve injury.Conclusion Extremity ischemic preconditioning has protective effect on pul-monary function in the patients undergoing pulmonary lobectomy,which may be related to down-reg-ulation of TLR4 expression in monocytes of blood and inhibition of the systemic inflammatory re-sponse.

Objective To investigate plasma NGF expression in adriamycin induced rat heart failure (HF) model .Methods Twenty‐five Wistar rats were randomly divided into the CHF group (n=15 ,adriamycin 4 mg/kg ,by intraperitoneal injection ,for 6 weeks) and normal control group (NC group ,n=10) ,after successful model construction ,the 6‐week observation was continuously conducted .The body mass and plasma NGF expression were detected once per 2 weeks .Results After 6 weeks later ,the body mass in the CHF group was significantly reduced ,the difference was statistically significant (P<0 .05) ,LVEDV and LVESV were sig‐nificantly increased ,while LVEF was declined obviously (P<0 .05) ,the NGF expression amount was significantly decreased com‐pared with the control group ,the differences were statistically significant (P<0 .05);the NGF expression amount was gradually re‐duced with the time extension of disease course(P<0 .05) .Conclusion Intraperitoneal injection of adriamycin can successfully in‐duce heart failure model in Wistar rats ,moreover NGF may be closely associated with HF .

OBJECTIVETo observe the impacts of electroacupuncture (EA) at Zusanli (ST 36) and Feishu (BL 13) applied 30 min before the operation till the end of the operation on the postoperative inflammatory reaction and pulmonary complications in the senile patients after radical resection of pulmonary carcinoma.

METHODSEighty senile patients of pulmonary carcinoma were selected and randomized into an observation group and a control group, 40 cases in each one. In the observation group, EA stimulation at Zusanli (ST 36) and Feishu (BL 13) was used 30 min before the operation till the end of the operation. In the control group, electric stimulation was not used. Separately, before operation (T, basic state), 12 h after operation (T) and 24 h after operation (T), blood sample was collected from the central vein. The concentrations of plasma tumor necrosis factor-ɑ (TNF-ɑ) and interleukin-10 (IL-10) were detected. Additionally, the radial arterial blood sample was collected at the above time points; oxygen partial pressure (PaO) was determined; pulmonary alveoli-arterial partial pressure of oxygen (PDO) and oxygenation index (OI) were calculated. The pulmonary complication in the two days after operation was recorded.

RESULTSCompared with the control group, in the observation group, at Tand T, TNF-ɑ concentration and PDOwere lower (all<0.05); plasma IL-10 concentration and OI were higher (all<0.05). In the observation group, the incidences of postoperative pneumonia and acute pulmonary injury were lower than those in the control group (both<0.05).

CONCLUSIONSEA reduces the postoperative inflammatory reaction in the senile patients with radical resection of pulmonary carcinoma and decreases the postoperative pulmonary complicattizen.