Objective:To optimize the chitosan flocculation purification process of Zhigancao decoction. Methods:With the total content of ginsenoside Rg1 , Re and Rb1 and the impurity removal rate as the evaluation indices, an orthogonal test was used to investi-gate the effects of concentration of solution, amount of chitosan and standing temperature on the purification process. Results:The op-timum conditions of chitosan flocculation clarification process of Zhigancao decoction were as follows:the concentration of extraction so-lution was 0. 25 g·ml-1 , the standing temperature was 25℃, and the amount of chitosan was 12% . Conclusion:The optimum con-ditions of purification process are promising with high impurity removal rate and active ingredients retention rate, which provide refer-ence for the dosage form reformation of Zhigancao decoction.

Objective:To explore the effects of holly leaf extract on blood lipid regulating function and blood rheology in hyperlipi-demia rats to provide experimental basis for the development and application of holly leaf. Methods:The rats were fed with high-fat di-et for 4 weeks to establish the model of hyperlipidemia. The rats were randomly divided into the control group, the model group, the positive group, high-dose group (0.60 g·mL-1), medium-dose group (0.30 g·mL-1) and low-dose group (0.15 g·mL-1) of Holly leaf with gastric perfusion for 9 weeks. TC, TG, HDL-C, LDL-C, AST, ALT, SOD and MDA in serum of the rats were deter-mined, and various indices of hemorheology were measured. Results:When compared with the model group, holly leaf extract at dif-ferent doses could decrease the content of TC, TG, LDL-C, ALT, AST, MDA and apo B in varying degrees, and increase the content of HDL-C, the SOD activity and lipoprotein levels in varying degrees at the end of the gastric perfusion, and the effects of high-dose of holly leaf were the most significant (P<0. 05,P<0. 01). Holly leaf extract at different doses could decrease plasma viscosity, whole blood viscosity, erythrocyte aggregation index and hematocrit, and increase red blood cell deformability index, and the effects of high-dose group of holly leaf were the most significant (P<0. 05,P<0. 01). Conclusion:Holly leaf has obvious regulating effects on blood lipid function and blood rheology in hyperlipemia rats, which can reduce blood lipid levels.

Aim To evaluate the effects of gavage with lactococcus lactis(L.Lactis) recombinant heme oxyge-nase-1(HO-1) gene on alleviation of intestinal inflammation and protection of the intestinal mucosa in rats with endotoxemia.Methods A model of rats with endotoxemia was produced in 24 healthy SD male rats.The rats were randomly divided into the L.Lactis recombinant HO-1(LL-HO-1)gene group(HO-1 group,n=8),L.Lactis group(LL group,n=8) and glutamine group(Glu group,n=8).The rats received 1 ml(5?1010 CFU?ml-1)?d-1LL-HO-1,1 ml(5?1010 CFU?ml-1)?d-1 L.Lactis or 1 g?kg-1?d-1 glutamine for four days before LPS intraperitoneal injecion.Twenty hours after LPS injection,the full-thickness distal ileum was harvested.The mortality,myeloperoxidase(MPO) activity,the pathological changes,the contents of HO-1,tumor necrosis factor-?(TNF-?) and interleukin-10(IL-10) in the intestine were determined.Results When the results of HO-1 group was compared with those of LL group,the survival rate was significantly elevated in the former(P

Objective To compare the effects oflightwand(LW) and direct laryngoscope(DL) in elderly patients undergoing surgery on hemodynamics and stress responses.Methods Forty elderly patients with ASA grade Ⅰ-Ⅱ grade who selective surgery were divided into LW group and DL group by random digits table method,each group with 20 cases.Systolic blood pressure (SBP),diastolic blood pressure (DBP),heart rate(HR) were recorded respectively before intubation,30 seconds and 5 minutes after intubation.Extracted patients with peripheral arterial 3 ml at the same time,measured by radioimmunoassay epinephrine and norepinephrine levels.Results The SBP,DBP,HR,epinephrine,norepinephrine 30 seconds after intubation in LW group were significantly lower than those in DL group [(140.50 ± 21.91) mm Hg(1 mm Hg =0.133 kPa) vs.(167.95 ± 17.85) mm Hg,(67.80 ± 6.76) mm Hg vs.(84.15 ±9.89) mm Hg,(85.10 ± 11.76) times/min vs.(96.30 ± 13.72) times/min,(49.47 ± 8.09) ng/L vs.(61.25 ± 9.43) ng/L,(171.27 ± 17.11) ng/L vs.(187.40 ± 16.13) ng/L,P ＜ 0.01].The SBP,epinephrine,norepinephrine 5 minutes after intubation in LW group were significantly lower than those in DL group [(120.75 ± 17.12) mm Hg vs.(136.65 ± 15.43) mm Hg,(39.80 ±7.38) ng/L vs.(47.63 ±8.48) ng/L,(155.93 ± 17.11) ng/L vs.(172.76 ± 13.45) ng/L,P ＜ 0.01].Conclusion Both LW and DL can be well done in elderly patients intubation,but LW can reduce stress responses during the intubation.

Objective To investigate the distribution of five indicators of hepatitis B(HBV-M)and its relationship with liver function parameters and HBV-DNA load in patients with chronic HBV infection in different stages.Methods The serum samples were collected from 456 patients infected with HBV.The HBV-M,liver functional parameters and HBV-DNA level were quantita-tively detected.According to the stages of disease,the patients were divided into 3 groups including chronic hepatitis B group(inclu-ding mild subgroup,moderate subgroup and severe subgroup),liver cirrhosis group(including compensatory subgroup,decompen-sated subgroup)and hepatocellular carcinoma goup.Results The ratio of HBsAg,HBeAg,HBcAb positive pattern(135 pattern) and HBsAg,HBeAb,HBcAb positive pattern(145 pattern)in the three groups were statistically different(P <0.05).In each chron-ic hepatitis B subgroup,both ALT and AST levels of 135 pattern were significantly higher than the other two patterns(P <0.05). In each liver cirrhosis subgroup and hepatocellular carcinoma group,there were statistically significant differences in ALB and TBIL levels between the three patterns(P <0.05 ).In each group,the HBV-DNA level of 135 positive pattern were significantly higher than the other two patterns(P <0.05).Conclusion With the advancement of chronic hepatitis B,there is a downtrend in the ratio of 135 pattern and increasing trend in the ratio of 145 pattern.If the stage of hepatitis B is discriminated,ALT,AST,ALB TBIL and HBV-DNA level were closely related to HBV-M pattern.

Objective To compare the safety and efficacy of intravenous thrombolysis for patients with acute cerebral infarction in a second-grand class-A hospital and a third-grand class-A hospital. Methods Twenty-one consecutive patients with cerebral infarction treated with alteplase for intravenous thrombolysis were enrolled in a second-grand class-A hospital (Fengtai Hospital,Beijing)prospectively from January 2012 to December 2013 as the study group,and 65 patients in a third-grand class-A teaching hospital (Xuanwu Hospital,Capital Medical University,Beijing)admitted at the same period for intravenous thrombolysis were used as a control group. The differences of efficacy and safety of intravenous thrombolysis in patients of both groups were compared. The primary outcome measures were Barthel Index (BI)at day14 after onset and the modified Rankin Scale (mRS)scores at discharge. The main safety indicator was the incidence of serious adverse events (SAEs)after thrombolysis (symptomatic intracranial hemorrhage and death). Results (1 )In the primary outcome measures,the proportions of mRS≤2 at discharge in the study group and the control group were 71. 4%(n=15)and 58. 5%(n=38)respectively. At day 14 after thrombolysis,the proportions of BI ≥60 were 61. 9%(n=13)and 64. 6%(n=42)respectively. There were no significant differences between the two groups (P>0. 05). (2)The incidences of the primary serious adverse events were 4. 8%(n=1)and 6. 2%(n=4). There was no significant difference (P>0. 05). Other secondary outcome measures,such as the early reperfusion rate,recanalization rate,and the proportion of neurological improvement at day 14 after thrombolysis and the overall incidence of cerebral hemorrhage had no significant differences. The case referral proportion (9. 5%,n=2)of the study group had a trend of lowering than the control group (27. 7%,n=18)P=0. 09. (3)The out-hospital time delay, in-hospital time delay,and overall time delay of the study group were less than those of the control group, and the mean time was 75 ± 33 vs. 102 ± 50 min,and 72 ± 41 vs. 111 ± 38 min,147 ± 41 vs. 212 ± 47 min. There were significant differences (P<0. 01). Conclusion The second-grand hospital selected by our study can relatively safely and effectively perform intravenous thrombolysis for acute cerebral infarction with alteplase. Moreover,the intravenous thrombolysis of the second-grand hospitals may reduce the case referral ratio and visiting time.

The transcription activity of ectogenic human carcinoembryonic antigen (CEA) promoter in lung adenocarcinoma cells A549 was investigated for the further gene-targeting therapy. The reporter gene green fluorescent protein (GFP) driven by CEA promoter and human cytomegalovirus (CMV) promoter were relatively constructed and named plasmid pCEA-EGFP and pCMV-GFP respectively. The intensity of fluorescence was detected by fluorescence microscope and flow cytometry analysis after the pCEA-GFP and pSNAV-GFP plasmids were transfected into A549 cells through liposome respectively. The results showed (4.08+/-0.63) % of the A549 cells transfected with pCEA-AFP plasmid expressed, significantly lower than that of the A549 cells transfected with pCMV-GFP [(43.27+/-3.54) %]. It was suggested that ectogenic human CEA promoter in lung adenocarcinoma cells A549 was weakly expressed. The distinct specificity of CEA promoter in CEA high expression cells was regarded as a tool in selective gene therapy, but the transcription activity of ectogenic human CEA promoter was needed to increase in the future.

Objective To evaluate the effect of methylene blue (MB) on hydrogen peroxide (H2O2)-induced apoptosis in macrophages through mitochondria-dependent pathway in mice.Methods Mouse peritoneal macrophage line RAW264.7 cells were cultured in DMEM culture medium containing 10％ fetal bovine serum.Cells were divided into 6 groups (n =24 each) using a random number table method:control group (group C),H2O2 group,prophylactic different concentrations of MB groups (MB1,2 groups) and therapeutic different concentrations of MB groups (MB3.4 groups).H2O2 50 μmol/L was added to the culture medium in group H2O2.MB was added to the culture medium with the final concentrations of 0.1 μmol/L (in MB1 and MB3 groups) and 1.0 μmol/L (in MB2 and MB4 groups) at 30 min before adding H2O2 in MB1.2 groups and 30 min after adding H2O2 in MB3.4 groups.At 24 h of culture or incubation in each group,the cell survival rate was measured by methyl thiazolyl tetrazolium assay,the activity of reactive oxygen species (ROS) in cells was determined with the fluorescent probe,the lactate dehydrogenase (LDH) activity in supernatant was detected by spectrophotometry,the activity of superoxide dismutase (SOD) in cells was detected by colorimetric method,mitochondrial membrane potential (MMP) was measured using rhodamine 123 staining,the content of ATP was determined by an ATP bioluminescent method,the expression of pro-caspase-3 and spliceosomes P20 protein and P 18 protein was detected by Western blot,and cell apoptosis was detected using flow cytometry.Results Compared with group C,the cell survival rate,SOD activity and contents of MMP and ATP were significantly decreased,the ROS activity and activity of LDH in supernatant were increased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was up-regulated,and early and late apoptosis rates were increased in the other five groups (P＜0.05).Compared with group H2O2,the cell survival rate,SOD activity and contents of MMP and ATP were significantly increased,the ROS activity and activity of LDH in supernatant were decreased,the expression of pro-caspase-3 and spliceosomes P20 protein and P18 protein was down-regulated,and early and late apoptosis rates were decreased in MB1-4 groups (P＜0.05).Compared with group MB1,the cell survival rate was significantly decreased,and the expression of caspase-3 spliceosome P 18 was down-regulated in group MB2,and the cell survival rate and SOD activity were significantly decreased,and the activity of ROS was increased in group MB3 (P＜0.05).Compared with group MB4,the expression of caspase-3 spliceosome P 18 was significantly down-regulated,early and late apoptosis rates were decreased,and the activity of ROS was increased in group MB2,and the activity of ROS was significantly increased in group MB3 (P＜0.05).Conclusion The mechanism by which MB attenuates H2O2-induced oxidative damage to macrophages is related to inhibiting cell apoptosis in macrophages through mitochondria-dependent pathway in mice.

Objective:To investigate the preoperative evaluation, safety and clinical efficacy of intravascular recanalization for patients with symptomatic non-acute middle cerebral artery occlusion .Methods:Twenty-six patients with symptomatic non-acute middle cerebral artery occlusion admitted between January 2018 to June 2021 were retrospectively analyzed. The clinical symptoms, cerebral perfusion, occlusive site, occlusive length, collateral circulation and distal capillary were evaluated. There were 17 cases accorded with intravascular recanalization. The cerebral blood flow , cerebral blood volume, MTT, peak time and scores of 17 patients before and after revascularization were compared by paired t test.Results:Endovascular recanalization was performed in 17 patients and extracranial-intracranial artery bypass grafting was performed in 9 patients. Among the 17 cases, there were 7 cases of hemiplegia of contralateral limbs, 6 cases of recurrent cerebral infarction, 4 cases of paroxysmal contralateral limb numbness or weakness, 3 cases of cognitive dysfunction and 3 cases of headache. The cerebral perfusion of the occluded side of the middle cerebral artery in 17 cases was significantly lower than that of the healthy side. One patient failed to pass through the occlusion with micro guide wire, and gave up the operation after repeated attempts. The occluded vessels were successfully recanalized in 16 cases (thrombolytic grade of cerebral infarction=2b-3), and recanalized in 16 cases, including 9 cases of simple balloon dilatation and 7 cases of stent implantation. Three months after the operation, the National Institute of Health Stroke Scale (NIHSS) score of 17 patients decreased from (9.57±2.32) to (3.75±1.42). The cerebral blood flow, cerebral blood volume, mean transit time and peak time were also significantly improved compared with those before the operation（ t value was 9.08，5.54，4.26，8.56，6.00，respectively，all P<0.001）. The Montreal Cognitive Assessment (MoCA) score of cognitive function was (22.70±1.70) before the operation, and there months after operation (26.30±2.30) points ( t=6.66, P<0.001) were statistically significant. After operation, CT cerebral angiography or whole cerebral angiography showed that 1 case was occluded, 15 cases had good recanalization, no intracranial hemorrhage occurred during the perioperative period, and no new stroke occurred during the 6-month follow-up period. Conclusion:For patients with symptomatic non acute middle cerebral artery occlusion, the patients who may benefit from multi-dimensional evaluation are selected for intravascular recanalization treatment. There are few complications and high recanalization rate. The short-term follow-up clinical effect is good, but the long-term effect needs to be further observed.

Background/Aims: Previous studies have shown that diet and physical activity can influence constipation. However, the combined effect of diet and physical activity on constipation remains unclear. Methods: Constipation was defined based on stool consistency and frequency, while overall diet quality was assessed using Healthy Eating Index (HEI)-2015 scores. Participants were categorized into low (metabolic equivalent [MET]-min/wk < 500) and high physical activitygroups (MET-min/wk ≥ 500). The association between diet and constipation across physical activity groups was analyzed using surveylogistic regression and restricted cubic splines. Results: Higher HEI-2015 scores were associated with reduced constipation risk in the high physical activity group when constipation was defined by stool consistency (odds ratio [OR], 0.98; 95% confidence interval [CI], 0.97-0.99). However, in the low physical activity group, increased HEI-2015 scores did not significantly affect constipation risk (OR, 1.01; 95% CI, 0.97-1.05). Similar results were found when constipation was defined based on stool frequency. In the high physical activity group, increased HEI-2015 scores were significantly associated with a reduced constipation risk (OR, 0.96; 95% CI, 0.94-0.98). Conversely, in the low physical activity group, increased HEI-2015 scores did not affect the risk of constipation (OR, 0.96; 95% CI, 0.90-1.03). Conclusions Our findings suggest that a higher HEI-2015 score is negatively associated with constipation among individuals with high physical activity levels but not among those with low physical activity levels. This association was consistent when different definitions of constipation were used. These results highlight the importance of combining healthy diet with regular physical activity to alleviate constipation.