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 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 establish an animal model of chronic systemic inflammation with long-term high expression of circulating IL-6 by introducing exogenous IL-6 gene transfer vector.Methods:Recombinant murine IL-6-encoding adeno-associated virus (AAV-IL-6) was constructed. Twenty-one 24-week-old male C57BL/6J mice were randomly divided into three groups with seven in each group: AAV-IL-6 group, vector control (AAV-ctrl) group and blank control group. At 0, 8 and 16 weeks of intervention, the mice in the three groups were injected with AAV-IL-6 (100 μl 0.5×10 10 vp/ml), unloaded AAV (100 μl 0.5×10 10 vp/ml) and the same volume of saline in the tail vein, respectively. IL-6 levels in mouse serum were measured by ELISA. The general condition of mice was observed and blood routine tests were performed. Changes in blood biochemical parameters and C-reactive protein (CRP) levels were detected. At the end of 24-week intervention, the mice were sacrificed and the myocardium, liver, spleen, quadriceps femoris, knee joint and middle femur were taken for HE staining. Results:At 4, 8, 16 and 24 weeks after intervention, serum IL-6 levels were (75.41-169.28) pg/ml in the AAV-IL-6 group, while in the two control groups, the levels were below the lower limit of detection (7.8 pg/ml). At 24 weeks after intervention, the body weight of mice in the AAV-IL-6 group was significantly lower than that of mice in the two control groups; the neutrophil counts and CRP level in the AAV-IL-6 group were higher than those in the two control groups, while the levels of albumin, creatinine, triglyceride and cholesterol were lower than those in the two control groups. There were no differences in the aforementioned parameters between the two control groups. Compared with the blank control group, both AAV-IL-6 and AAV-ctrl groups showed increased lymphocyte counts. All mice had normal liver and kidney functions at the end of intervention. Histopathological findings indicated that the mice in the AAV-IL-6 group had focal infiltration of lymphocytes in the central venous area of the liver and around the myocardial and the skeletal muscle fibers, diffuse infiltration of multinucleated giant cells in the spleen, atrophic skeletal muscle, disorganized growth plate, reduced chondrocyte hypertrophic zone, thinner bone cortex and trabecular, and reduced osteoid. There were no histopathological changes in mice of the two control groups.Conclusions:Repeated tail vein injection of AAV-IL-6 could achieve long-term high expression of circulating IL-6 in mice, which manifested the phenotype of chronic systemic inflammation in preliminary detection and provided a safe, effective and simply accessible animal model for related studies.

Objective:To explore the trans-membrane signaling mechanism of interleukin-6 (IL-6)-induced osteogenic differentiation and calcification of human umbilical artery smooth muscle cells (HUASMCs).Methods:HUASMCs were primarily cultured in vitro and were stimulated with IL-6, IL-6+solutable IL-6 receptor (sIL-6R), IL-6+sIL-6R+solutable gp130 (sgp130), or vehicle (blank control). Alizarin red and Von Kossa staining were used for detecting cell calcification, Western blot was used to test the protein expression of tissue-nonspecific alkaline phosphatase (TNAP), osteopontin (OPN), bone morphogenetic protein-2 (BMP-2) and Runt related transcription factor 2 (Runx2), and immunofluorescence was used to examine the mIL-6R expression of HUASMCs. The comparison of measurement date between the two groups was conducted by t-test. The comparison of measurement date between multiple groups was conducted by one-way analysis of variance (ANOVA). Results:The intensity severity of calcification stain was IL-6+sIL-6R group >IL-6+sIL-6R+sgp130 group>IL-6 group=blank control. After stimulated for 12 hours, the TNAP expression in blank control, IL-6 group, IL-6+sIL-6R group, IL-6+sIL-6R+sgp130 group were (0.44±0.08), (0.52±0.14), (0.84±0.16) and (0.55±0.10) respectively ( F=290.96, P<0.001). After stimulated for 3 days, the OPN expression in blank control, IL-6 group, IL-6+sIL-6R group, IL-6+sIL-6R+sgp130 group were (0.61±0.84), (0.95±0.16), (1.65±0.24) and (0.99±0.10) respectively ( F=507.72, P<0.001). After stimulated for 12 hours, the BMP-2 expression in blank control, IL-6 group, IL-6+sIL-6R group, IL-6+sIL-6R+sgp130 group were (0.77±0.05), (1.69±0.16), (2.81±0.26) and (0.57±0.12) respectively ( F=959.09, P<0.001). After stimulated for 3 days, the Runx2 expression in blank control, IL-6 group, IL-6+sIL-6R group,IL-6+sIL-6R+sgp130 group were (0.57±0.03) , (0.92±0.10), (1.31±0.13) and (0.66±0.06) respectively ( F=1141.27, P<0.001). Comparing with Jurkat cells (positive control) and CEM cells (negative control), HUASMCs limited expressed mIL-6R. Conclusion:IL-6 may induce HUASMCs osteogenic differentiation and calcification mainly via the sIL-6R-mediated trans-signaling pathway.

Objective:To explore early-onset gout and related risk factors in Shandong Province, and provide decision-making information on prevention.Methods:Data from electronic medical records and face-to face interview were collected from 8 393 patients with gout who first visited the gout clinic of the Affiliated Hospital of Qingdao University from September 2016 to December 2021. Data included demographics, comorbidity and biochemical examinations. The dynamic changes of onset age from 2002 to 2021 were statistically analyzed. The clinical characteristics and related risk factors of patients with early-onset and late-onset gout were statistically analyzed.Results:The age of onset of gout decreased significantly from 2002 to 2021. Compared with 2002, the average age of onset in 2021 decreased by 2.3 years [(41.9±10.6 vs 39.6±14.0) years]. The median age of onset decreased by 3 years in 2012-2021 compared with 2002-2011(37 vs 40 years, P<0.001). The proportion of gout patients with onset age<40 years old increased significantly, from 45.1% in 2002 to 57.8%, and increased by 12.7% in 20 years( P<0.001). The constituent ratios of 20-29 years old group( Ptrend<0.001) and≤19 years old group( Ptrend=0.011) increased by 9.3%( P<0.001) and 4.2%( P=0.002) over 20 years, which was the highest increase among all age groups with onset age<40 years old. Multivariate stepwise linear regression analysis showed that positive family history, blood uric acid level, metabolic syndrome and smoking were independent risk factors for early onset of gout. Conclusion:The age of gout onset in tends to be younger. The increase of the proportion of patients younger than 30 years old is probably the key factor leading to the early-onset gout in Shandong Province. Early and effective intervention on the risk factors related to early-onset gout is essential to prevent the early-onset gout as well as to reduce the prevalence of gout and complications.

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 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.

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.