Objective:To evaluate the effect of hydrogen on lipopolysaccharide (LPS) and nigericin-induced pyroptosis in macrophages and the role of long non-coding RNA (lncRNA) nuclear-enriched abundant transcript 1 (NEAT1).Methods:Human monocyte-derived macrophages THP-1 cells were cultured in vitro and divided into 4 groups ( n=25 each) using a random number table method: control group (group C), LPS and nigericin group (group LN), hydrogen-rich medium+ LPS and nigericin group (group H+ LN), and lentiviral transfection+ hydrogen-rich medium+ LPS-nigericin group (group LV+ H+ LN). THP-1 cells were cultured in the common culture medium for 24 h in group C. LPS at a final concentration of 100 ng/ml and nigericin 10 μmol/L were added to the culture medium, and the cells were incubated for 24 h in group LN. In group H+ LN, the culture medium was replaced with 0.6 mmol/L hydrogen-rich medium, then LPS at a final concentration of 100 ng/ml and nigericin 10 μmol/L were immediately added, and the cells were incubated for 24 h. In group LV+ H+ LN, THP-1 cells over-expressing NEAT1 stably after being transfected with lentivirus were used, then LPS at a final concentration of 100 ng/ml and nigericin 10 μmol/L were immediately added, and the cells were incubated for 24 h. Cell viability was detected by CCK-8 assay. Lactic dehydrogenase (LDH) release was assessed by colorimetric method. The amount of LDH released was measured by colorimetry. The concentrations of interleukin-1beta (IL-1β) and IL-18 in culture medium were measured by enzyme-linked immunosorbent assay. The pyroptotic rate was detected by flow cytometry. The expression of nucleotide-binding oligomerization domain-like receptor-containing pyrin domain 3 (NLRP3), apoptosis-associated speck-like protein (ASC), caspase-1 and gasdermin D (GSDMD) was detected by Western blot. The expression of NEAT1 gene was determined by quantitative real-time polymerase chain reaction. Results:Compared with group C, the cell viability was significantly decreased, the amount of LDH released, concentrations of IL-1β and IL-18, and pyroptotic rate were increased, and the expression of NEAT1 gene, NLRP3, ASC, caspase-1 and GSDMD was up-regulated in group LN ( P<0.05). Compared with group LN, the cell viability was significantly increased, the amount of LDH released, concentrations of IL-1β and IL-18, and pyroptotic rate were decreased, and the expression of NEAT1 gene, NLRP3, ASC, caspase-1 and GSDMD was down-regulated in group H+ LN ( P<0.05). Compared with group H+ LN, the cell viability was significantly decreased, the amount of LDH released, concentrations of IL-1β and IL-18, and pyroptotic rate were increased, and the expression of NEAT1 gene, NLRP3, ASC, caspase-1 and GSDMD was up-regulated in group LV+ H+ LN ( P<0.05). Conclusions:Hydrogen can ameliorate LPS and nigericin-induced pyroptosis in macrophages, and the mechanism may be associated with down-regulating the expression of lncRNA NEAT1.

Objective:To investigate the correlation between plasma microRNA (miR)-122, miR-33a and the severity of coronary artery disease in patients with type 2 diabetes mellitus (T2DM) and coronary heart disease.Methods:The clinical data of 196 patients with T2DM from January 2019 to October 2021 in Xuzhou First People′s Hospital were retrospectively analyzed. Among them, 81 cases were complicated with coronary heart disease (combined group), 115 cases were not complicated with coronary heart disease (control group). The plasma levels of miR-122 and miR-33a were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction, the plasma level of N-terminal B-type natriuretic peptide precursor (NT-proBNP) was detected by enzyme-linked immunosorbent assay. In combined group, the number of coronary artery lesions was determined according to the results of coronary angiography, and Gensini score was evaluated. Linear regression model was used to analyze the relationship between plasma miR-122, miR-33a and NT-proBNP levels with the incidence of coronary heart disease in patients with T2DM. Receiver operating characteristic (ROC) curve was used to analyze the plasma miR-122 and miR-33a in predicting efficiency of coronary heart disease in patients with T2DM. In combined group, Spearman correlation method was used to analyze the relationship between plasma miR-122, miR-33a and the number of coronary artery lesions, and Pearson correlation method was used to analyze the relationship between plasma miR-122, miR-33a and plasma NT proBNP, Gensini score.Results:The plasma miR-122, miR-33a and NT-proBNP in combined group were significantly higher than those in control group: 5.76 ± 1.35 vs. 1.18 ± 0.33, 1.39 ± 0.37 vs. 0.65 ± 0.11 and (786.87 ± 156.39) ng/L vs. (103.45 ± 19.27) ng/L respectively, and there were statistical differences ( P<0.01). Linear regression result showed that plasma miR-122, miR-33a, and NT-proBNP were positive correlation with occurrence of coronary heart disease in patients with T2DM ( P<0.01); ROC curve analysis result showed that the area under curve of plasma miR-122, miR-33a and combination in predicting coronary heart disease in patients with T2DM were 0.816, 0.845 and 0.912 respectively (95% CI 0.744 to 0.865, 0.768 to 0.892 and 0.836 to 0.967). Coronary angiography result showed that there were 46 cases of single vessel lesions, 25 cases of double vessel lesions and 10 cases of three vessel lesions. The plasma miR-122, miR-33a, NT-proBNP and Gensini score in patients with three vessel lesions were significantly higher than those in patients with double vessel lesions and patients with single vessel lesions: 6.52 ± 0.96 vs. 4.95 ± 0.85 and 3.74 ± 0.52, 1.45 ± 0.31 vs. 1.06 ± 0.25 and 0.81 ± 0.13, (829.78 ± 62.59) ng/L vs. (627.48 ± 47.12) and (502.64 ± 38.24) ng/L, (63.89 ± 12.71) scores vs. (42.18 ± 6.03) and (22.36 ± 2.41) scores, the indexes in patients with double vessel lesions were significantly higher than those in patients with single vessel lesions, and there were statistical differences ( P<0.05). In combined group, Spearman correlation analysis result showed that the plasma miR-122 and miR-33a were positive correlation with the number of coronary artery lesions ( r = 0.879 and 0.825, P<0.05); Pearson correlation analysis result showed that the plasma miR-122 and miR-33a were positive correlation with the plasma NT-proBNP and Gensini score (miR-122: r = 0.896 and 0.788, miR-33a: r = 0.871 and 0.765; P<0.05). Conclusions:The plasma levels of miR-122 and miR-33a are related to the occurrence of coronary heart disease and severity of coronary artery disease in patients with T2DM, which may be used to guide the prevention and treatment of coronary heart disease in patients with T2DM.

Objective To evaluate the relationship between the change rate of regional cerebral ox-ygen saturation ( rSO2 ) and early postoperative cognitive function during laparoscopic surgery in elderly pa-tients. Methods Fifty American Society of Anesthesiologists physical status Ⅰ-Ⅱ patients, aged 65-80 yr, scheduled for elective laparoscopic radical resection for colorectal cancer under general anesthesia, were enrolled in this study. rSO2 was recorded at 5 min before anesthesia induction ( T0 ) , 5 min after tra-cheal intubation ( T1 ) , 5 min after changing position ( T2 ) , 1 h after changing position ( T3 ) , 5 min after stopping pneumoperitoneum ( T4 ) , and 5 min after changing to supine position at the end of pneumoperito-neum ( T5 ) . The change rate of rSO2 was calculated at T3 . Mini-Mental State Examination scores were per-formed at 1 day before surgery and 3 days after surgery. The patients were divided into cognitive disorder group and non-cognitive disorder group. Results Compared with non-cognitive disorder group, rSO2 was significantly increased at T2-5 , and the change rate of rSO2 was enlarged in cognitive disorder group ( P<0. 05) . The change rate of intraoperative rSO2 was negatively correlated with Mini-Mental State Examination scores at 3 days after surgery (r=-0. 516, P<0. 01). Conclusion The increased change rate of rSO2 during laparoscopic surgery may be related to the development of early postoperative cognitive dysfunction in elderly patients.

Objective To evaluate the effect of oxycodone pretreatment on oxygen-glucose deprivation and restoration (OGD/R)-induced injury to small intestinal epithelial cells of rats and the role of different opioid receptors.Methods IEC-6 cells were divided into 5 groups (n=15 each) using a random number table:control group (group C),group OGD/R,oxycodone group (group O),μ opioid receptor antagonist CTOP plus oxycodone group (group CTOP+O) and κ opioid receptor antagonist BNI plus oxycodone group (group BNI+O).Cells were cultured for 8 h in normal culture atmosphere in group C.Cells were subjected to OGD for 4 h followed by restoration of oxygen-glucose supply in normal culture medium for 4 h in OGD/R,O,CTOP+O and BNI+O groups.Oxycodone at a final concentration of 1 μg/ml was added at 5 min before OGD/R in O,CTOP+O and BNI+O groups.CTOP and BNI at a final concentration of 5 μmol/L were added at 10 min before OGD/R in group CTOP+O and group BNI+O,respectively.Five holes in each group were selected at 8 h of OGD/R for determination of the cell viability (by MTT assay),lactic dehydrogenase (LDH) release rate (by colorimetry) and levels of tumor necrosis factor-alpha (TNF-α),interleukin-1beta (IL-1β) and high-mobility group box 1 protein (HMGB1) in supernatant (by enzyme-linked immunosorbent assay).Results Compared with group C,the cell viability was significantly decreased,and the LDH release rate and levels of TNF-α,IL-1β and HMGB1 were increased in OGD/R,O,CTOP+Oand BNI+ O groups (P＜ 0.05).Compared with group OGD/R,the cell viability was significantly increased,and the LDH release rate and levels of TNF-o,IL-1β and HMGB1 were decreased in group O (P＜0.05).Compared with group O,the cell viability was significantly decreased,and the LDH release rate and levels of TNF-α,IL-1β and HMGB1 were increased in CTOP+O and BNI+O groups (P＜0.05).Conclusion Oxycodone pretreatment can mitigate OGD/R-induced injury to small intestinal epithelial cells of rats and the mechanism is related to activating μ and κ opioid receptors.

Objective To evaluate the effect of oxycodone on intestinal ischemia-reperfusion injury in rats and the role of autophagy.Methods Twenty-four healthy adult male Sprague-Dawley rats,aged 6-9 weeks,weighing 180-220 g,were divided into 4 groups (n =6 each) using a random number table method:sham group (group S),intestinal I/R group (group I/R),oxycodone group (group O) and oxycodone plus autophagy inhibitor 3-methyladenine (3-MA) group (group O+3-MA).Intestinal I/R was induced via clamping the superior mesenteric artery for 1 h,followed by 2-h reperfusion in all the groups except for group S.Oxycodone 0.5 mg/kg was injected via caudal vein at 15 min before ischemia in group O and group O+3-MA.3-MA 30 mg/kg was intraperitoneally injected at 10 min before ischemia in group O+3-MA.Rats were gavaged with fluorescein-isothiocyanate-conjugated dextran (FITC-dextran) immediately before ischemia.Blood samples were collected from the cardiac apex at 2 h of reperfusion to detect the level of serum FITC-dextran.Blood samples were collected from the cardiac apex at 2 h of reperfusion to measure the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in serum by enzyme-linked immunosorbent assay.Small intestinal tissues were obtained at 2 h of reperfusion for examination of the pathological changes and for determination of the expression of occludin,Beclin-1 and microtubule-associated protein 1 light chain 3 (LC3) (by Western blot).Intestinal damage was assessed and scored according to Chiu.The ratio of LC3 Ⅱ expression to LC3 Ⅰ expression (LC3 Ⅱ / Ⅰ) was calculated.Results Compared with group S,the Chiu's score,levels of serum FITC-dextran,TNF-α and IL-1β and LC3 Ⅱ/Ⅰ ratio were significantly increased,the expression of Beclin-1 was up-regulated,and the expression of occludin was down-regulated in group I/R (P＜0.05).Compared with group I/R,the Chiu's score and levels of serum FITC-dextran,TNF-α and IL-1β were significantly decreased,LC3 Ⅱ / Ⅰ ratio was increased,and the expression of occludin and Beclin-1 was up-regulated in group O (P＜0.05),and no significant change was found in the parameters mentioned above in group O+ 3-MA (P＞0.05).Compared with group O,the Chiu's score and levels of serum FITC-dextran,TNF-α and IL-1β were significantly increased,LC3 Ⅱ / Ⅰ ratio was decreased,and the expression of occludin and Beclin-1 was down-regulated in group O + 3-MA (P＜ 0.05).Conclusion Oxycodone can ameliorate intestinal I/R injury,and the mechanism may be related to enhancing autophagy in intestinal tissues of rats.

Objective To evaluate the efficacy of preoperative transversus abdominis plane (TAP) block for improving analgesia after radical resection of colorectal cancer in elderly patients.Methods Fiftysix American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients of both sexes,aged 75-86 yr,weighing 52-78 kg,scheduled for elective radical resection of colorectal cancer under general anesthesia,were divided into 2 groups (n=28 each) using a random number table:TAP block group (group T) and control group (group C).After anesthesia induction,ultrasound-guided bilateral TAP block was performed,and 0.40％ ropivacaine 25 ml was injected into each side in group T.Both groups received patientcontrolled intravenous analgesia (PCIA) with fentanyl after surgery.PCIA solution contained fentanyl 600 μg and azasetron 10 mg in 100 ml of normal saline.The PCIA pump was set up with a 2 ml bolus dose,a 15 min lockout interval and background infusion at a rate of 2 ml/h,and visual analogue scale score was maintained≤ 3.When the visual analogue scale score＞3,tramadol 50-100 mg was intravenously injected as rescue analgesic.The number of patients requiring rescue analgesic,consumption of fentanyl during PCIA and the number of successfully delivered doses were recorded within 24 h after surgery.The adverse reactions such as hematoma at the puncture site,nausea and vomiting,respiratory depression,chest tightness and pruritus were recorded.Results Compared with group C,the number of patients requiring rescue analgesic was significantly decreased,and the consumption of fentanyl and the number of successfully delivered doses during PCIA were decreased in group T (P＜0.05).No hematoma was found at the puncture site in group T.No respiratory depression,chest tightness or pruritus was found in the two groups,and there were no significant differences in the incidence of nausea and vomiting between the two groups (P＞0.05).Conclusion Preoperative TAP block can reduce the consumption of fentanyl and enhance the efficacy of analgesia after radical resection of colorectal cancer in elderly patients.

Objective To evaluate the effect of 6% hydroxyethyl starch 130∕0. 4 on acute kidney injury in elderly patients in a prospective, multicenter, randomized, double?blind, controlled clinical tri?al. Methods A total of 120 patients of both sexes, aged 65-82 yr, weighing 56-83 kg, of American So?ciety of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective orthopaedics and hernia surgery, were divided into either hydroxyethyl starch group ( group HES) or lactated Ringer′s solution group ( group LR) , with 60 patients in each group. Hydroxyethyl starch and lactated Ringer′s solution were infused intra?venously at a rate of 7. 5 ml∕kg during 1st hour of surgery in HES and LR groups, respectively. Lactated Ringer′s solution was then infused at a rate of 5 ml∕kg starting from 2nd hour of surgery until the end of sur?gery in both groups. Before surgery, at the end of surgery and at 1, 3 and 5 days after surgery, blood sam?ples and urine specimens were collected for determination of the concentrations of neutrophil gelatinase?asso?ciated lipocalin, interleukin?18 (IL?18), plasma creatinine, urine β2 microglobulin and urine albumin.
The estimated glomerular filtration rate was calculated. Results The level of urine IL?18 was significantly higher at each time point after surgery than before surgery and immediately after the end of surgery ( P<0.05) . There were no significant differences between the two groups in the levels of urine IL?18, plasma creatinine, plasma and urine neutrophil gelatinase?associated lipocalin, plasma IL?18, urineβ2 microglob?ulin and urine albumin and estimated glomerular filtration rate at each time point ( P>0.05) . Conclusion Compared with lactated Ringer′s solution, 6% hydroxyethyl starch 130∕0.4 does not aggravate acute kidney injury in elderly patients.

Objective To evaluate the effects of propofol and sevoflurane anesthesia on cognitive function and amyloid beta protein ( Aβ) deposition in hippocampi of aged mice. Methods Thirty?six SAMP8 mice, aged 6 months, weighing 29-32 g, were randomly assigned into 4 groups ( n=9 each) using a random number table: control group ( group C ) , propofol anesthesia group ( group P ) , sevoflurane anesthesia group (group S) and propofol plus sevoflurane anesthesia group (group PS). In group P, propofol 140 mg∕kg was injected intraperitoneally, when righting reflex occurred, additional propofol 70 mg∕kg was given, and when it occurred again, additional propofol 40 mg∕kg was given. Group S continuously inhaled 1% sevoflurane for 120 min. Group PS continuously inhaled 2% sevoflurane for 120 min, and when righting reflex occurred, additional propofol 40 mg∕kg was given. Anesthesia was maintained for 120 min in P, S and PS groups. Before anesthesia and at 7, 14 and 28 days after anesthesia, Morris water maze test was performed, and the escape latency was recorded. Hippocampi were obtained to determine the expression of Aβ using immuno?histochemistry. Results Compared with group C, the escape latency was significantly prolonged at 7 days after anesthesia, and the expression of Aβwas up?regulated at 7, 14 and 28 days after anesthesia in group S, and no significant change was found in the parameters mentioned above in P and PS groups. Compared with the value at 7 days after anesthesia, the expression of Aβ was significantly down?regulated at 14 and 28 days after anesthesia in group S, and no significant change was found in the expression of Aβ at 14 and 28 days after anesthesia in C, P and PS groups. Conclusion Although sevoflurane anesthesia promotes Aβ deposition in hippocampi, it only causes short?term cognitive dysfunction, however, anesthesia with propofol or with propofol in combination with sevoflurane produces no influence in aged mice.

Objective To evaluate the role of histone methylation in the spinal cord in maintenance of neuropathic pain (NP) in rats.Methods Sixty healthy male Wistar rats, weighing 200-250 g, aged 6-8 weeks, were randomly divided into 5 groups (n =12 each) using a random number table: control group (C group), sham operation group (S group), NP group, chaetocin group (CH group), and dimethyl sulfoxide (DMSO) group.NP was induced by chronic constriction injury (CCI) to the sciatic nerve in NP, CH and DMSO groups.The sciatic nerve was only exposed in group S.H3K9 methylation methyltransferase inhibitor chaetocin 50 μg was injected intrathecally in group CH, and the equal volume of DMSO was given instead in group DMSO for 4 consecutive days, starting from 2nd day after CCI.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before CCI (T0),before administration on 2nd day after CCI (T1), and at 5, 7, 9 and 14 days after CCI (T2-5).Six rats in each group were sacrificed after measurement of pain threshold at T3 and T5, and the lumbar segment of the spinal cord was removed for detection of the methylated H3K9 expression by immunohistochemistry.Results Compared with C group, no significant change was found in the MWT, TWL and methylated H3K9 expressionat each time point in S group (P＞0.05) , the MWT was significantly decreased, and the TWL was shortened at T1-5, and the methylated H3K9 expression was up-regulated at T3 and T5 in NP and DMSO groups, and the MWT was significantly decreased, and the TWL was shortened at T1 in CH group (P＜ 0.05).Compared with NP group, the MWT was significantly increased, and the TWL was prolonged at T2-5, and the methylated H3K9 expression was down-regulated at T3 and T5 in CH group (P＜0.05) , and no significant change was found in the MWT, TWL and methylated H3K9 expression at each time point in DMSO group (P＞0.05).Conclusion Histone methylation in the spinal cord is involved in the maintenance of NP in rats.

Objective To evaluate the effects of different head-down tilt (HDT) angles on respiratory mechanics and cerebral circulation during laparoscopic radical resection of rectal carcinoma in the patients.Methods Thirty patients of both sexes,aged 40-64 yr,of ASA physical status Ⅰ or Ⅱ,with body mass index ＜ 30 kg/m2,scheduled for elective laparoscopic radical resection of rectal carcinoma,were included in the study.Anesthesia was induced with midazolam,fentanyl,propofol and cis-atracurium and maintained with targetcontrolled infusion of propofol and remifentanil and cis-atracurium when needed.At 5 min after supine position,5 min after 30° HDT,5 min after 35° HDT,5 min after 40° HDT,and 30 min after pneumoperitoneum (T1-5),airway pressure (Paw),peak airway pressure (PIP),lung compliance (Cpat) and jugular bulb blood pressure (JBP) were recorded.Blood samples were taken from the radial artery and jugular bulb for blood gas analysis and arterial partial pressure of oxygen (PaO2),partial pressure of carbon dioxide in artery (PaCO2),arterial oxygen saturation (SaO2),jugular bulb oxygen partial pressure (PjvO2),and jugular bulb venous saturation (SjvO2) were recorded at each time point mentioned above.Results Paw,PIP,MAP,PaCO2,JBP,PjvO2 and SjvO2 were significantly increased as compared with those obtained when the patients were in supine position,increased gradually with the increasing HDT angles,and decreased to the level obtained when the patients were in supine position at 30 min after pneumoperitoneum.Cpat and PaO2 were significantly decreased as compared with those obtained when the patients were in supine position decreased gradually with the decreasing HDT angles,and increased to the level obtained when the patients were in supine position at 30 min after pneumoperitoneum.Conclusion HDT-induced effects on respiratory mechanics and cerebral circulation increase with increasing angles during laparoscopic radical resection of rectal carcinoma; thus the optimum HDT angle is smaller than 40° for the patients undergoing this kind of surgery.