Objective:To discuss the effects of monocyte chemoattractant protein-1(MCP-1)on the migration and invasion of lung cancer A549 cells,and to clarify the mechanisms.Methods:Immunohistochemistry method was used to detect the expression of MCP-1 protein in 80 cases of non-small cell lung cancer(NSCLC)and adjacent normal lung tissues.The human lung cancer A549 cells were cultured in vitro.The MCP-1-small interfering RNA(siRNA)experiment was divided into blank group,negative control group(si-NC group),MCP-1-siRNA-1 group,and MCP-1-siRNA-2 group.The MCP-1 over-expression experiment was divided into control group,empty vector control group(OE-NC,transfected with MCP-1 over-expression empty vector),over-expression MCP-1 group(OE-MCP-1 group,transfected with MCP-1 over-expression plasmid),over-expression MCP-1+extracellular regulated protein kinase(ERK)pathway inhibitor PD98059 group(OE-MCP-1+PD98059 group,co-transfected with MCP-1 over-expression plasmid and PD98059),and PD98059 group(transfected with PD98059).The MCP-1 siRNA and plasmids were transfected into the lung cancer A549 cells;Western blotting method was used to verify the transfection efficiencies of the cells in various groups;the migration rate and the number of invasion cells in various groups were observed by wound healing assay and Transwell chamber assay,respectively;Western blotting method was also used to detect the expression levels of phosphorylated ERK(p-ERK),total ERK(t-ERK),and epithelial-mesenchymal transition(EMT)-related proteins in the A549 cells in various groups.Results:Compared with adjacent tissue,the positive expression rate of MCP-1 protein in NSCLC tissue was significantly increased(P<0.05),and the expression level of MCP-1 protein was related to TNM stage and lymph node metastasis(P<0.05).Compared with si-NC group,the expression level of MCP-1 protein in the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups was significantly decreased(P<0.01).Compared with control group and OE-NC group,the expression level of MCP-1 protein in the cells in OE-MCP-1 group was significantly increased(P<0.01).The wound healing assay results showed that compared with si-NC group,the migration rate of the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups were significantly decreased(P<0.01).Compared with OE-NC group,the migration rate of the cells in OE-MCP-1 group was significantly increased(P<0.01);compared with OE-MCP-1 group,the migration rate of the cells in OE-MCP-1+PD98059 group was significantly decreased(P<0.01).Compared with OE-MCP-1+PD98059 group,the migration rate of the cells in PD98059 group was significantly decreased(P<0.01).The Transwell chamber assay results showed that compared with si-NC group,the number of invasion cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups was significantly decreased(P<0.01).Compared with OE-NC group,the number of invasion cells in OE-MCP-1 group was significantly increased(P<0.01);compared with OE-MCP-1 group,the number of invasion cells in OE-MCP-1+PD98059 group was significantly decreased(P<0.01);compared with OE-MCP-1+PD98059 group,the number of invasion cells in PD98059 group was significantly decreased(P<0.01).The Western blotting results showed that compared with si-NC group,the expression levels of p-ERK,Vimentin,and N-cadherin protein in the cells in MCP-1-siRNA-1 and MCP-1-siRNA-2 groups were significantly decreased(P<0.05 or P<0.01),and the expression level of E-cadherin proteins was significantly increased(P<0.01).Compared with OE-NC group,the expression levels of p-ERK,Vimentin,and N-cadherin proteins in the cells in OE-MCP-1 group were significantly increased(P<0.01),and the expression level of E-cadherin protein was significantly decreased(P<0.01).Compared with OE-MCP-1 group,the expression levels of p-ERK,Vimentin,and N-cadherins proteins in the OE-MCP-1+PD98059 group were significantly decreased(P<0.01),and the expression level of E-cadherin protein was significantly increased(P<0.05).Compared with OE-MCP-1+PD98059 group,the expression levels of p-ERK,Vimentin,and N-cadherin proteins in the cells in PD98059 group were significantly decreased(P<0.05 or P<0.01),and the expression level of E-cadherin protein was increased(P<0.01).Conclusion:MCP-1 protein can upregulate the expression of EMT-related proteins in the lung cancer A549 cells,and promote the migration and invasion of the lung cancer A549 cells;its mechanism may be related to the activation of the ERK signaling pathway.

Objective To explore the optimal ratio of Baitong decoction based on efficacy,clarify its spectrum-effect relationship,and identify its potential quality markers.Methods An ulcerative colitis(UC)model in mice was established using dextran sulfate sodium.The efficacy of Baitong decoction with varying drug ratios was assessed by evaluating the apparent score,pathological score and inflammatory factor changes of UC in each group of experimental animals.The fingerprints of Baitong decoction with different ratios were established by high performance liquid chromatography(HPLC),and the relationship between the content of each substance and its efficacy was analyzed by partial least squares regression to determine the potential quality markers of Baitong decoction.Results Baitong decoction was most effective in relieving ulcerative colitis when the mass ratio of Fuzi,Ganjiang and Congbai was 1∶2∶2.The fingerprinting identified 14 common peaks across 7 ratios,with 9 peaks were found to be associated with the remission of ulcerative colitis by partial least squares regression analysis.Conclusion The optimal ratio of Fuzi,Ganjiang and Congbai for treating UC is 1∶2∶2.The spectrum-effect relationship analysis suggested that the quality markers of Baitong decoction may be the substances represented by peak 2(benzoylaconine),3,5,6,8(mesaconine),9(aconitine),10(hypaconitine),13(10-gingerol)and 14.

Objective:To evaluate the relationship between breast cancer resistance protein (BCRP)-blood brain barrier (BBB) and dexmedetomidine-induced improvement in postoperative cognitive function in patients with mild hyperbilirubinemia.Methods:This was a prospective study. Ninety patients of both sexes with mild hyperbilirubinemia caused by choledocholithiasis, aged 55-69 yr, with body mass index of 22-28 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, with preoperative Mini-Mental State Examination (MMSE) scores≥20, scheduled for elective cholecystectomy, exploratory choledocholithotomy and T-tube drainage from December 2022 to August 2023 in the Third Central Hospital of Tianjin, were divided into 2 groups( n=45 each) using a random number table method: control group (C group) and dexmedetomindine group (D group). After induction of anesthesia, dexmedetomidine was intravenously infused at a loading dose of 0.5 μg/kg over 10 min, followed by an infusion of 0.6 μg·kg -1·h -1 until the end of operation in group D, and the equal volume of normal saline was given instead in group C. At 2 days before operation and 1, 3, 5, 7 and 14 days after operation, the cognitive function was assessed using MMSE and Montreal Cognitive Assessment, and the serum BCRP concentration, concentrations of cognitive function serological indicators (serum S100β, β amyloid 42, malondialdehyde), and concentrations of BBB serological indicators (serum glial fibrillary acidic protein, thrombospondin 1, claudin 5) were determined using enzyme-linked immunosorbent assay. Results:Compared with group C, MMSE and Montreal Cognitive Assessment scores were significantly increased, the incidence of cognitive impairment was decreased, the serum concentrations of S100β, β amyloid 42 and malondialdehyde were decreased, the serum concentrations of BCRP were increased, and the serum concentrations of glial fibrillary acidic protein, claudin-5 and thrombospondin 1 were decreased in group D ( P<0.05). Conclusions:The mechanism by which dexmedetomidine improves postoperative cognitive function may be related to up-regulating BCRP levels and improving BBB function in patients with mild hyperbilirubinemia.

Objective:To identify the risk factors for postoperative cognitive dysfunction (POCD) and develop the prediction model in elderly patients undergoing lumbar surgery under general anesthesia.Methods:The elderly patients undergoing elective lumbar surgery under general anesthesia in our hospital from July 2021 to July 2022 were enrolled. Cognitive function was assessed at 7 days after surgery using Mini-Mental State Examination and Montreal Cognitive Assessment. When the decrease in both scales≥ 1 standard deviation, the patients were considered as having POCD. The patients were divided into POCD group and non-POCD group according to whether POCD developed. The propensity score matching was used to balance the confounding bias between two groups. The multivariate logistic regression analysis was used to identify the risk factors for POCD. The prediction model was constructed, and a nomogram was drawn for visualization of the model. The receiver operating characteristic curve, calibration plot and decision curve analysis (DCA) were drawn to evaluate the differentiation, consistency and clinical validity of the model, respectively.Results:A total of 159 patients were enrolled in this study, and the incidence of POCD was 31.4%. There were statistically significant differences in the ratio of intraoperative blood transfusion, cumulative time of hypotension, total infusion volume and operation time between two groups ( n=32 each) after propensity score matching ( P<0.05). The results of multivariate logistic regression showed that age, educational levels, diabetes mellitus, previous two or more operations under general anesthesia, APTT and cumulative time of hypotension were independent risk factors for POCD in elderly patients undergoing lumbar surgery under general anesthesia ( P<0.05). A model was developed based on the risk factors mentioned above: LogitP=-15.878+ 0.263 × Age (years) - 0.122 × Educational Level (years)+ 1.601 × Diabetes Mellitus+ 1.468 × History of General Anesthesia for 2 or more times+ 0.608 × Cumulative Time of Hypotension(min) - 0.140 × APTT (s). The area under the receiver operating characteristic curve was 0.930 (95% CI 0.887-0.973), the sensitivity was 0.920, specificity was 0.798 and Youden index was 0.718. After visualizing the model via nomogram, the model was verified by Hosmer-Lemeshow test, P=0.403, C index was 0.930, and corrected C index was 0.914. Conclusions:Age, educational levels, diabetes mellitus, previous multiple operations under general anesthesia, APTT and cumulative time of hypotension are independent risk factors for POCD in elderly patients undergoing lumbar surgery under general anesthesia, and the established risk prediction model can effectively predict the occurrence of POCD in elderly patients undergoing lumbar surgery under general anesthesia.

Objective:To prepare and preliminary verify dezocine polylactic acid-glycolic acid block copolymer (PLGA) microspheres.Methods:Preparation of dezocine PLGA microspheres Dezocine 120 mg, PLGA 0.1 g and the solubilizing additive poloxamer 0.1 g were dispersed in tetrahydrofuran solvent to form an organic phase solution. Sodium chloride and polyethylene glycol were dissolved in water for injection to form an inner aqueous phase solution and an outer aqueous phase solution. After the organic phase solution 20 ml was mixed with the inner aqueous phase solution 20 ml to form a water/oil colostrum, the water/oil colostrum was added to the outer aqueous phase solution to form a water/oil/water multiple emulsion, which was fully mixed with lyophilized powder protective agent and freeze-dried to prepare dezocine PLGA microspheres. Verification Eighteen clean-grade healthy male Sprague-Dawley rats, aged 10-12 weeks, weighing 220-260 g, were divided into 3 groups ( n=6 each) by a random number table method: control group (group C), dezocine ordinary preparation group (group D 1) and dezocine PLGA microspheres group (group D 2). Normal saline, dezocine injectio (dose 1 mg) and dezocine PLGA microsphere injectio (dose 0.2 μg) 0.2 ml were intramuscularly injected in C, D 1 and D 2 groups, respectively. The concentrations of dezocine in plasma were measured at 30 min and 1, 2, 3, 4, 5, 6, 7 and 8 h after administration, and thermal paw withdrawal latency was measured at T 1-T 3, T 5 and T 9. Tissues from the injection site were obtained on day 7 after intramuscular injection, and the inflammatory response was observed after HE staining. Results:Compared with group C, the thermal paw withdrawal latency was significantly prolonged at T 1-T 3 in group D 1 and at T 1-T 3, T 5 and T 9 in group D 2 ( P<0.05). Compared with group D 1, the thermal paw withdrawal latency was significantly prolonged at T 5 and T 9, and the plasma concentrations of dezocine were increased at T 6-T 9 in group D 2 ( P<0.05). Compared with the values at T 2, the plasma concentrations of dezocine were significantly decreased at T 4-T 9 in group D 1 ( P<0.05), and no significant change was found in the plasma concentrations of dezocine at T 3-T 9 in group D 2 ( P>0.05). On 7 days after injection, no inflammation was found in the local tissues in C, D 1 and D 2 groups, and no significant difference was found among the three groups. Conclusions:The sustained-release formulation of dezocine PLGA microspheres is successfully prepared, and it can maintain stable blood concentrations, effectively prolongs the action time of the drug and has significant sustained-release effect in rats.

Objective:To determine the median effective dose (ED 50) and the 95% effective dose (ED 95) of remifentanil inhibiting responses to endotracheal intubation without neuromuscular relaxant when combined with dexmedetomidine in patients undergoing thyroid surgery. Methods:American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of either sex, aged 18-64 yr, with body mass index of 18-28 kg/m 2, scheduled for elective thyroid surgery under intraoperative neuromonitoring, were enrolled in this study.Dexmedetomidine was intravenously injected in a loading dose of 0.8 μg/kg at 10 min before anesthesia induction.Anesthesia was induced by intravenously injecting midazolam 0.1 mg/kg, etomidate 0.4 mg/kg and the preset dose of remifentanil.The dose of remifentanil was determined using up-and-down sequential method.The initial dose was set at 3.7 μg/kg.The dose of remifentanil in the next case was determined according to whether responses to endotracheal intubation occurred, and the ratio between the two successive doses was 1.1.The ED 50, ED 95 and 95% confidence interval (CI) were calculated by Probit analysis. Results:when combined with dexmedetomidine for anesthesia induction, the ED 50 (95% CI) of remifentanil inhibiting responses to endotracheal intubation without neuromuscular relaxant was 3.39 (3.29-3.50) μg/kg, and the ED 95 (95% CI) was 3.52 (3.48-3.64) μg/kg. Conclusion:when combined with dexmedetomidine, the ED 50 of remifentanil inhibiting responses to endotracheal intubation without neuromuscular relaxant is 3.39 μg/kg, and the ED 95 is 3.52 μg/kg.

Objective:To evaluate the effects of CD34 + cell transplantation on radiation-induced brain injury (RIBI) and the relationship with the activity of astrocytes in rats. Methods:Healthy adult male Sprague-Dawley rats, weighing 210-230 g, were divided into 3 groups ( n=36 each) using a random number table method: control group (C group), RIBI group, and CD34 + group.RIBI model was established by computed tomography (CT) scanning in anesthetized rats.Another 6 rats were selected, and CD34 + cells were eluted by flow cytometry and labeled with BrdU.CD34 + cells were transplanted at day 7 after establishing the model.Brain tissues were obtained at 7, 14 and 28 days after establishing the model in C and RIBI groups and at 14 and 28 days after establishing the model in CD34 + group for determination of Evans blue (EB) extravasation ratio and expression of GFAP (by immuno-histochemistry). Results:Compared with group C, the EB extravasation ratio was significantly increased after establishing the model, and the expression of GFAP was up-regulated in group RIBI ( P<0.05), and no significant change was found in EB extravasation ratio after establishing the model in group CD34 + ( P>0.05). Compared with group RIBI, the EB extravasation ratio was significantly decreased after establishing the model, and the expression of GFAP was down-regulated in group CD34 + ( P<0.05). Conclusion:CD34 + cell transplantation can reduce RIBI, and the mechanism may be related to inhibiting the activity of astrocytes in rats.

Objective To observe the aortic calcification level in patients with rheumatoid arthritis (RA),and to analyze the relationships between aortic calcification and some RA disease related presentations.Methods RA patients (RA group) were all in-patients consecutively recruited from the Department of Rheumatology in one single tertiary hospital,and healthy subjects (control group) were individuals for check-up from the same hospital at the same time.Subjects with long-term smoking and drinking history,diabetes,hypertension,coronary heart disease,cancer,active or chronic infection,other autoimmune diseases and liver or kidney dysfunction were excluded in both groups.The aortic calcification scores (including ascending aorta,arcus aorta and aorta thoracica) were obtained automatically by 256-slice spiral CT scanner using the Heart Beat-CS program.Statistical package from Soci-science (SPSS) 17.0 software was used for data analysis.Student's t test,Mann-Whitney U test,Spearman test and x2 test were used.Results One hundred RA patients and 60 healthy subjects were selected,and there were no differences of age [(53±10) vs (51 ±8),t=1.031,P=0.304) and gender compositions [male 40(40％) vs 25(41％),x2=0.430,P=0.869) between the two groups.The aortic calcification score in the RA group was higher than that in the control group [19.4(3.3,190.0) vs 2.1 (1.9,18.0),U=1 579.5,P＜0.01].In RA group,the calcification score was positively correlated with age (r=0.729,P＜0.01),course of disease (r=0.227,P=0.023),C-reactive protein (CRP) (r=0.229,P=0.022),total cholesterol (TC) (r=0.220,P=0.028) and low density lipoprotein cholesterol (LDL-C) (r=0.224,P=0.014),but not related with treatment duration,number of tender joints and swollen joints,erythrocyte sedimentation rate,rheumatoid factor,anti-CCP antibody,DAS-28 (CRP),DAS-28 (ESR),triglyceride (TG) and high density lipoprotein cholesterol (HDL-C).The aortic calcification was also positively correlated with age in control group (r=0.465,P＜0.01),but not related with TC,TG,HDL-C,LDL-C.Conclusion RA patients have more severe aortic calcification than the matched general population.Aortic calcification degree is related to disease course,CRP,TC and LDL-C,which indicates that chronic systemic inflammation is essential to aortic calcification in RA.

Objective To investigate the correlation between the severity of alcoholic fatty liver disease and the amount of fat in the abdominal cavity and the serum inflammatory factor IL-18 and IL-8. Methods From October 2016 to October 2017,one hundred and twenty patients with AFLD in the First Hospital of Hebei Medical University were divided into light,medium,heavy groups according to the severity of fatty lesions by color Doppler Ultrasound. There were 40 mild patients,50 moderate patients and 30 severe patients. Forty healthy subjects were selected as controls. All the participants underwent CT scanning. The intra-abdominal fat area (VAT),abdominal subcutaneous fat area (SAT) and total abdominal fat area (TA) were measured. The liver function was measured by biochemical analyzer and enzyme-linked immunoassay (ELISA). (ELSIA) IL-18 was detected and IL-8 was detected by radioimmunoassay. Results The VAT of the healthy control group and the mild,medium and severe AFLD group were (70. 28±10. 19),(114. 38 ± 9. 97),(146. 73±10. 19),(163. 38±12. 69) cm2. The TA of the healthy control group and the mild, medium and severe AFLD group were ( 256. 72± 34. 56),( 332. 19 ± 33. 28),( 387. 49± 32. 28),( 478. 19 ±31. 02) cm2. The SAT of the healthy control group and the light,medium and severe AFLD group were (156. 23±28. 19),(203. 43±27. 12),(246. 19±26. 89),(271. 19 ±27. 94) cm2,respectively. Aspartate aminotransferase (AST) of the healthy control group and the mild,medium and severe AFLD group were (18. 50±1. 12),(23. 50±1. 21),(25. 50±1. 24),(29. 50± 1. 43) U/L. Alanine aminotransferase (ALT) of the healthy control group and the light, medium and severe AFLD group were ( 18. 50 ± 2. 14), ( 26. 50 ±2. 22),(35. 50±2. 34),(38. 50±2. 11) U/L. γ-glutamyltransferaseof the healthy control group and the light,medium and severe AFLD group were ( 16. 50 ± 2. 11), ( 32. 50 ± 2. 23), ( 47. 50 ± 2. 31), ( 48. 00 ±2. 43) U/L,respectively. Compared with the healthy control group,VAT,TA,SAT,AST,ALT andγ-GT in the light,medium and heavy AFLD group showed statistically significant differences ( P＜0. 05) . Compared with the mild AFLD group, VAT, TA, SAT, AST, ALT and γ-GT in the medium and heavy AFLD group showed statistically significant differences ( P＜0. 05) . Compared with the moderate AFLD group,the VAT, TA,SAT, AST, ALT, and γ-GT of the severe AFLD group showed statistically significant differences ( P＜0. 05). The data of the three AFLD groups showed that the concentration of all indicators were increasing as the severity of fat deepened. IL-18 of the healthy control group and the light,medium and severe AFLD group were (45. 67±4. 51),(52. 18±5. 09),(59. 87±4. 98),(64. 18±5. 12) ng/L; IL-8 of the healthy control group and the light, medium and severe AFLD group were ( 78. 92 ± 5. 07), ( 115. 62 ± 4. 89), ( 223. 76 ± 6. 78),(286. 42±7. 02) g/L. Compared with every group,IL-18 and IL-8 of light,medium and severe AFLD group showed statistically significant differences (F＝1035. 67,2. 93×105,P＜0. 001); compared with mild AFLD group,IL-18 and IL-8 of medium and heavy group showed statistically significant differences;compared with moderate AFLD group,IL-18 and IL-8 of severe group AFLD showed statistically significant differences ( P＜0. 001) . The levels of inflammatory factors IL-18 and IL-8 increased with the severity of steatosis. The severity of AFLD was significantly positively correlated with VAT,TA,SAT,IL-18 and IL-8 ( r 0. 415(P＜0. 001), 0. 435 ( P＜0. 001), 0. 512 ( P＜0. 001), 0. 274 ( P＜0. 001 ), 0. 689 ( P ＜0. 001). Conclusion Fat control is an important measure to prevent AFLD. IL-18 and IL-8 can reflect the severity of liver injury in AFLD and have important significance in judging prognosis.

Objective: To evaluate the effect of dexmedetomidine on postoperative cognitive function in the patients with mild hyperbilirubinemia caused by choledocholithiasis. Methods: One hundred and twenty patients of both sexes with mild hyperbilirubinemia (serum total bilirubin levels 21-170 μmol/L) caused by choledocholithiasis, aged 51-63 yr, with body mass index of 20-28 kg/m², of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, with preoperative Mini-Mental State Examination (MMSE) scores≥20, scheduled for elective cholecystectomy and choledocholithotomy, were divided into 3 groups (n=40 each) using a random number table method: control group (C group) and dexmedetomindine 0.4 μg·kg^-1·h^-1 group (D1 group) and dexmedetomindine 0.6 μg·kg^-1·h^-1 group (D2 group). After induction of anesthesia, dexmedetomidine was intravenously infused for 10 min in a loading dose of 0.5 μg/kg, followed by an infusion of 0.4 and 0.6 μg·kg^-1·h^-1 until the end of operation in D1 and D2 groups, respectively.The equal volume of normal saline was given instead in group C. with preoperative scores≥20, MMSE and Montreal Cognitive Assessment (MoCA) were used to assess the cognitive function at 1 day before operation (T₀) and 1, 3, 5 and 7 days after operation (T_1-4). The occurrence of cognitive dysfunction within 7 days after operation was recorded.Venous blood samples were collected at the time points mentioned above, and the plasma concentrations of β-amyloid (Aβ) 42 were determined by enzyme-linked immunosorbent assay. Results: Compared with group C, MoCA scores were significantly increased at T₁ in group D₁, and MMSE scores at T₁ and MoCA scores at T₁ and T₂ were significantly increased, and the plasma concentrations of Aβ42 were decreased at T_2-4 in group D2, and the incidence of cognitive dysfunction was significantly decreased in D1 and D2 groups (P<0.05). Compared with group D1, MoCA scores were significantly increased at T₁, and the plasma concentrations of Aβ42 were decreased at T_2-4 in group D2 (P<0.05). Conclusion Dexmedetomidine can improve postoperative cognitive function, and intravenous infusion at a rate of 0.6 μg·kg^-1 · h^-1 provides better efficacy for the patients with mild hyperbilirubinemia caused by choledocholithiasis.