Objective To investigate the correlation of serum angiopoietin-1(Ang-1),angiopoietin-2(Ang-2),and Ang-1/Ang-2 ratio with HBA DNA and alanine aminotransferase(ALT)in patients with chronic hepatitis B(CHB)or liver cirrhosis.Methods Clinical data and serum specimens were collected from 99 patients with CHB and 59 patients with liver cirrhosis who were admitted to Beijing YouAn Hospital,Capital Medical University,from March 2018 to October 2019,and 46 individuals who underwent physical examination were enrolled as control group.PCR was used to measure serum HBV DNA level,and ELISA was used to measure the serum levels of Ang-1 and Ang-2.The serum levels of Ang-1 and Ang-2 and Ang-1/Ang-2 ratio were compared between groups.The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups,and the Bonferroni method was used for further comparison between two groups;the Spearman correlation analysis was used to investigate the correlation of Ang-1,Ang-2,and Ang-1/Ang-2 ratio with HBV DNA and ALT.Results Compared with the control group,the CHB group and the liver cirrhosis group had a significant reduction in the level of Ang-1(479.0 pg/mL and 208.4 pg/mL vs 671.0 pg/mL,both P＜0.05),and compared with the CHB group,the liver cirrhosis group had a significant reduction in the level of Ang-1(P＜0.001).Compared with the control group,the CHB group and the liver cirrhosis group had a significant increase in the level of Ang-2(286.1 pg/mL and 438.4 pg/mL vs 198.0 pg/mL,both P＜0.001),and compared with the CHB group,the liver cirrhosis group had a significant increase in the level of Ang-2(P＜0.001).Compared with the control group,the CHB group and the liver cirrhosis group had a significant reduction in Ang-1/Ang-2 ratio(1.6 and 0.5 vs 3.4,both P＜0.001),and compared with the CHB group,the liver cirrhosis group had a significant reduction in Ang-1/Ang-2 ratio(P＜0.001).The Spearman correlation analysis showed that in the CHB group,Ang-1 was negatively correlated with HBV DNA and ALT(r=-0.400 and-0.394,both P＜0.001),Ang-2 was positively correlated with HBV DNA and ALT(r=0.365 and 0.351,both P＜0.001),and Ang-1/Ang-2 ratio was negatively correlated with HBV DNA and ALT(r=-0.463 and-0.473,both P＜0.001);in the liver cirrhosis group,Ang-1,Ang-2,and Ang-1/Ang-2 ratio had no correlation with HBV DNA or ALT(all P＞0.05).Conclusion There are significant changes in the serum levels of Ang-1 and Ang-2 and Ang-1/Ang-2 ratio in patients with CHB or liver cirrhosis,and Ang-1,Ang-2,and Ang-1/Ang-2 ratio reflects the degree of liver injury in patients with CHB to a certain extent.

[Objective] To determine the feasibility of preparing plasminogen (Pg) with Fraction Ⅲ precipitation (hereinafter referred to as FⅢ-P) from low-temperature ethanol process by full chromatography (hereinafter referred to as FⅢ-P process). [Methods] The FⅢ-P was diluted with dissolution buffer at different dilution times and stirring time. The potency and antigen concentration of Pg in dissolution sample were detected and the dissolution and clarification conditions were determined. Pre-treatment of loading sample and pre-experiment of affinity chromatography were carried out on the FⅢ-P dissolution sample to judge whether the loading sample had an impact on the chromatography by observing the performance of the affinity chromatography column and to evaluate whether the affinity chromatography could achieve the purpose of purifying Pg by detecting the plasma protein antigen concentration and Pg potency of the samples in the process. Two batches of FⅢ-P process were studied step by step, and the specific activity, steps and total recovery, and the output of Pg per ton of plasma were calculated. The feasibility of preparing Pg by FⅢ-P process was evaluated by comparing with the data of full chromatography process using plasma as raw material (hereinafter referred to as plasma process). [Results] The FⅢ-P was dissolved with 10 times of dissolution buffer, stirred for 1 hour, centrifuged at room temperature of 10 000×g for 15 minutes. The supernatant was first filtered with a screen, then clarified with an 8/0.8 μm filter, and finally filtered with a 0.45/0.2 μm filter and loaded. Pre-test showed that from clarification and filtration to Pg affinity chromatography, the step recovery of activity and antigen was 39.51% and 108.64%, respectively, the antigen concentration of Pg increased by 31.16 times and the activity increased by 11.39 times after affinity chromatography, which reaching the effect of affinity chromatography purification of Pg. The results of 2 batches of step-by-step scale-up FⅢ-P process showed that the total recoveries of antigen and activity from plasma to SP chromatography of FⅢ-P process were (45.76±1.10)% and (24.15±0.59)%, respectively, which had a total loss of about 1/3 of antigen and about 2/3 of activity compared to the plasma process. The Pg specific activity of SP chromatography eluent was (4.68±0.25) U/mg, which was about half of that of plasma process, but meeted the internal standard of > 4 U/mg. The output of Pg antigen per ton of plasma in the FⅢ-P process was 68.73% of that in the plasma process, and the output of Pg activity per ton of plasma in the plasma process was 29.82% of that in the plasma process, which basically achieved the purpose of waste utilization of FⅢ-P. [Conclusion] The technical route of preparing Pg from FⅢ-P by full chromatography is feasible.

Objective:To compare the test methods for aluminum content determination in vaccines by inductively coupled plasma-mass spectrometry(ICP-MS),inductively coupled plasma optical emission spectrometry(ICP-OES)and titration of the Chinese Pharmacopoeia 2020.Methods:The pre-treatment procedures,linearity,repeatability,accuracy,quantification limit,detection limit and sample determination results of the three methods were compared and analyzed by methodological verification and sample testing.Results:There was a good linear relationship of ICP-OES in the concentration range of 1-20μg·mL-1 aluminum content,r=0.9999.Aluminum content in 6 types of vaccines was in the range of 99％-104.6％,and RSDs were lower than 3％(n=9).There was a good linear relationship of ICP-MS in the concentration range of 2.5-80ng·mL-1(r=0.999).Aluminum content in 6 types of vaccines were in the range of 99.4％-108.9％,and RSDs were lower than 8％(n=9).There was no significant difference between the three methods in the determination of aluminum content in vaccines,and RSDs were lower than 10％.Conclusion:Both ICP-MS and ICP-OES methods can be used for the determination of alu-minum content in aluminum adjuvant vaccine.Both detection methods are simple,fast and accurate.ICP-OES has lower instrument costs and is easier to promote in the laboratory.

ObjectiveTo explore the effect of Babaodan （BBD） on the NOD-like receptor pyrin domain containing 3/cysteine aspartate-specific protease-3 （NLRP3/Caspase-1） pathway proteins in mice with acetaminophen （APAP）-induced acute liver injury. MethodC57BL/6 mice were randomly grouped， and BBD （75， 150， 300 mg·kg^-1， ig） was administered twice a day for three days. After 2 hours of the last administration， the mice were treated with APAP （400 mg·kg^-1， ip）， and the eyeballs were removed to collect blood after 14 hours. Then they were sacrificed by cervical dislocation for sample collection. Hematoxylin-eosin （HE） staining was used to observe the morphological changes of liver tissue cells， and biochemical methods were used to detect the activities of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， superoxide dismutase （SOD）， malondialdehyde （MDA） and myeloperoxidase （MPO） in serum of mice in each group. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to determine the mRNA expression of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β） and IL-6， and Western blot was performed to determine the protein expression of cyclooxygenase-2 （COX-2）， inducible nitric oxide synthase （iNOS）， NLRP3， Caspase-1 and IL-18 in the liver of mice. ResultCompared with the conditions in normal group， the hepatic lobule structure of mice in the model group was partially destroyed， and the hepatic sinusoids were dilated. And the expression levels of ALT and AST in serum， the protein levels of NLRP3， Caspase-1， iNOS， IL-18 and COX-2 and the mRNA levels of IL-1β， IL-6 and TNF-α were increased （P<0.05， P<0.01）. Compared with the model group， the administration groups had improvement in liver cell rupture and hepatic sinusoidal compression， and a dose-dependent decrease in the levels of ALT and AST in serum as well as the protein levels of NLRP3， Caspase-1， iNOS， IL-18 and COX-2 and the the mRNA levels of IL-1β， IL-6 and TNF-α in liver tissue （P<0.05， P<0.01）. ConclusionBBD can reduce APAP-induced acute liver injury in mice. The mechanism may be related to anti-oxidative stress， inhibition of NLRP3/Caspase-1 pathway， and decreased expression levels of IL-1β， IL-18， TNF-α and IL-6.

Objective:To analyze the distribution of blood lipids and apolipoprotein E ( ApoE) gene in patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province, and their relationship with ethnicity, gender, and age. Methods:A total of 102 patients with cardiovascular and cerebrovascular diseases who received treatment in Southern Central Hospital of Yunnan Province (The First People's Hospital of Honghe State) from May 2019 to June 2020 were included in this study. Their blood lipid and ApoE gene distributions were analyzed. The correlations between blood lipid and ApoE gene distribution and ethnicity, gender, and age were analyzed. Results:There was a significant difference in total cholesterol level between male and female patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province [(4.10 ± 1.27) mmol/L vs. (4.70 ± 1.83) mmol/L, t = 1.87, P = 0.048]. There was no significant difference in total cholesterol level among different ethnic groups and different ages ( P = 0.343, 1.000). There were no significant differences in low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglyceride levels among different ethnic groups, different sexes, and different ages ( P = 0.562, 0.125, 0.158; 0.884, 0.068, 0.681; 0.262, 0.367, 0.965). There were no significant differences in blood lipid levels among different ethnic groups, different sexes, and different ages ( P = 0.890, 0.336, 0.142). No E2/E4 gene was found in all patients. E2/E2 and E2/E3 genes of ApoE accounted for 16.67%, E3/E3 gene accounted for 70.59%, and E3/E4 and E4/E4 genes accounted for 12.74%. There was no significant difference in the distribution of ApoE gene among different ethnic groups, different sexes, and different ages ( χ2 = 0.13, 0.69, 0.44, P = 0.936, 0.429, 0.804). Conclusion:Female patients with cardiovascular and cerebrovascular diseases in Honghe Hani and Yi Autonomous Prefecture of Yunnan Province have higher total cholesterol levels than male patients. Low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, and triglyceride levels are not significantly correlated with ethnicity, sex, and age. Blood lipid abnormalities mainly manifest as low high-density lipoprotein cholesterol levels and high triglyceride levels. The E2/E4 gene has not been found in this region so far.

Objective:To analyze the distribution and clinical significance of cytochrome P 450 2C19 (CYP2C19) gene in patients with cardiovascular and cerebrovascular diseases in southern Yunnan. Methods:The data of 245 patients with cardiovascular and cerebrovascular diseases who received treatment in Southern Central Hospital of Yunnan Province between May 2019 and June 2020 were retrospectively analyzed. The distribution of CYP2C19 gene and its relationship with nationality, age, sex, blood lipids, hypertension, and diabetes were analyzed and compared between southern Yunnan and other regions.Results:The proportions of seven phenotypes of CYP2C19 gene *1/*17, *1/*1, *1/*2, *1/*3, *2/*2, *2/*3, *3/*3 in 245 patients were 2.86%, 38.37%, 39.18%, 5.31%, 9.39%, 4.08% and 0.82%, respectively. The proportions of individuals with superfast/ultrafast metabolism, fast metabolism, intermediate metabolism, and slow metabolism in 245 patients were 2.86%, 38.37%, 44.49%, and 14.29%, respectively. The frequency of polymorphisms in the CYP2C19 gene was consistent with the Hardy-Weinberg equilibrium ( P > 0.05), which was constant and representative. The Fisher test showed that the CYP2C19 gene distribution of patients with cardiovascular and cerebrovascular diseases in southern Yunnan was not greatly correlated with nationality, age, sex, underlying disease, blood lipids, and the types of cardiovascular and cerebrovascular diseases (all P > 0.05). There was a significant difference in CYP2C19 gene distribution in patients from southern Yunnan versus Dongguan, Jiangxi, Fujian, northern Sichuan, Chifeng, Xiamen, Shaanxi, and Kunming ( P < 0.001, < 0.001, 0.045, 0.008, 0.001, 0.005, < 0.001, 0.016). Conclusion:The distribution of CYP2C19 gene in patients with cardiovascular and cerebrovascular diseases in southern Yunnan is not obviously correlated with nationality, age, sex, underlying diseases, blood lipids, and the types of cardiovascular and cerebrovascular diseases. CYP2C19 gene distribution is related to regional distribution, which can guide personalized medication in different regions.

Objective:To analyze the distribution of solute carrier organic anion transporter family member 1b1 ( SLCO1B1) and apolipoprotein E ( ApoE) genes in a population from southern Yunnan. Methods:The data of 104 patients who received treatment in Southern Central Hospital of Yunnan Province (The First People's Hospital of Honghe State) between May 2019 and June 2020 were collected. The distribution of SLCO1B1 and ApoE genes and their relationship with nationality, sex, and age were analyzed and compared between different regions. Results:The percentage of patients carrying *1a/*1a, *1a/*1b, *1b/*1b, *1a/*15, *1b/*15, five phenotypes of SLCO1B1 gene, in the population from southern Yunnan was 4.81%, 32.69%, 42.31%, 12.50% and 7.69% respectively. Phenotypes *1a/*5, *5/*5, *5/*15 and *15/*15 were not detected. Normal metabolic phenotype of SLCO1B1 accounted for 79.81%, and intermediate metabolic phenotype of SLCO1B1 accounted for 20.19%. Weak metabolic phenotype was not detected. The percentage of patients carrying E2/E2, E2/E3, E3/E3, E3/E4, E4/E4, five phenotypes of ApoE gene in the population from southern Yunnan was 0.96%, 16.35%, 70.19%, 11.54% and 0.96% respectively. E2/E4 phenotype was not detected. The percentage of patients with ApoE protective phenotype, ApoE normal phenotype, and ApoE risk phenotype was 17.31%, 70.19% and 12.50% respectively. The observed polymorphism mutation frequency of SLCO1B1 and ApoE genes was consistent with the Hardy-Weinberg equilibrium ( P > 0.05), suggesting constancy and a population representation. The Fisher test showed that SLCO1B1 gene distribution differed significantly between ethnic minorities and Han nationality in southern Yunnan ( P = 0.013). There was no significant difference in SLCO1B1 gene distribution between different sexes and between different ages (all P > 0.05). There was no significant difference in ApoE gene distribution between ethnic minorities and Han nationality, between different sexes, and between different ages in the population from southern Yunnan (all P > 0.05). Conclusion:SLCO1B1 gene distribution is related to nationality in the population from southern Yunnan, but it is unrelated to sex and age. ApoE gene distribution is unrelated to nationality, sex and age.

Objective: To investigate the effects of tanshinone ⅡA on atherosclerosis plaque formation and adventitial mast cells activation in high-fat-diet induced Apo E^-/- mice model. Methods: Sixteen 8-week-old Apo E^-/-male mice and eight 8-week-old C57BL/6 male mice were randomly allocated into following group: the control group (C57BL/6 + carboxymethyl cellulose per gavage), the atherogenic group (Apo E^-/-+carboxymethyl cellulose per gavage) and the tanshinoneⅡA intervention group (Apo E^-/-+30 mg/kg tanshinone ⅡA per gavage). All three groups were fed with high-fat-diet for 26 weeks. Tanshinone ⅡA/carboxymethyl cellulose was applied by the method of gavage administration 6 weeks before execution. After 26 weeks, tumor necrosis factor-α (TNF-α) andinterleukin (IL)-6 levels in serum were assessed by ELISA. Carotid artery was removed, fixed with paraformaldehyde, embedded with paraffin and sectioned. Percentage of stenosis was evaluated on HE stained sections. Plaque progression was assessed by Movat staining. Toluidine blue staining was used to evaluate mast cells infiltration and activation. Immunochemistry staining was used to assess 5-HT, TNF-α and IL-6 expression. mRNA expression of mast cell marker Fcer1a in adventitial tissue was detected by real time-PCR. Results: After high-fat-diet for 26 weeks, the mice in the atherogenic group showed advanced atherosclerosis, tanshinoneⅡA intervention reduced the percentage of carotid artery stenosis caused by atherosclerotic plaque formation ((58.48±8.07)% vs. (80.31±4.08)%, P<0.05). Compared with the atherogenic group, tanshinone ⅡA intervention group had lower level of TNF-α ((12.39±1.62)pg/ml vs. (17.44±1.42)pg/ml) and IL-6 ((116.24±12.16)pg/ml vs. (166.05±19.09)pg/ml) in serum, lower TNF-α ((20 145±1 556) vs. (25 288±1 671)) and IL-6 ((25 688±1 604) vs. (35 286±4 198)) expression in adventitia (all P<0.05). TanshinoneⅡA intervention also decreased the number of mast cells infiltration and activation, reduced 5-HT expression and mast cell marker Fcer1a mRNA relative expression in adventitia (all P<0.05). Conclusions Tanshinone ⅡA could attenuate induced by high-fat-diet carotid artery atherosclerosis in Apo E^-/- mice. The protective effect of tanshinoneⅡA is probably mediated through reducing the number and activation percentage of mast cells, decreasing the release of inflammatory cytokines and inflammation of carotid artery in adventitia.

Objective To investigate the effects of tanshinoneⅡA on atherosclerosis plaque formation and adventitial mast cells activation in high?fat?diet induced Apo E?/?mice model. Methods Sixteen 8?week?old Apo E?/?male mice and eight 8?week?old C57BL/6 male mice were randomly allocated into following group: the control group (C57BL/6+carboxymethyl cellulose per gavage), the atherogenic group (Apo E?/?+carboxymethyl cellulose per gavage) and the tanshinoneⅡA intervention group (Apo E?/?+30 mg/kg tanshinoneⅡA per gavage). All three groups were fed with high?fat?diet for 26 weeks. TanshinoneⅡA/carboxymethyl cellulose was applied by the method of gavage administration 6 weeks before execution. After 26 weeks, tumor necrosis factor?α (TNF?α) andinterleukin (IL)?6 levels in serum were assessed by ELISA. Carotid artery was removed, fixed with paraformaldehyde, embedded with paraffin and sectioned. Percentage of stenosis was evaluated on HE stained sections. Plaque progression was assessed by Movat staining. Toluidine blue staining was used to evaluate mast cells infiltration and activation. Immunochemistry staining was used to assess 5?HT, TNF?α and IL?6 expression. mRNA expression of mast cell marker Fcer1a in adventitial tissue was detected by real time?PCR. Results After high?fat?diet for 26 weeks, the mice in the atherogenic group showed advanced atherosclerosis, tanshinoneⅡA intervention reduced the percentage of carotid artery stenosis caused by atherosclerotic plaque formation ((58.48±8.07)% vs. (80.31±4.08)%, P<0.05). Compared with the atherogenic group, tanshinoneⅡA intervention group had lower level of TNF?α ((12.39 ± 1.62)pg/ml vs. (17.44 ± 1.42)pg/ml) and IL?6 ((116.24 ± 12.16)pg/ml vs. (166.05 ± 19.09)pg/ml) in serum, lower TNF?α ((20 145±1 556) vs. (25 288±1 671)) and IL?6 ((25 688±1 604) vs. (35 286±4 198)) expression in adventitia (all P<0.05). TanshinoneⅡA intervention also decreased the number of mast cells infiltration and activation, reduced 5?HT expression and mast cell marker Fcer1a mRNA relative expression in adventitia (all P<0.05). Conclusions TanshinoneⅡA could attenuate induced by high?fat?diet carotid artery atherosclerosis in Apo E?/?mice. The protective effect of tanshinoneⅡA is probably mediated through reducing the number and activation percentage of mast cells, decreasing the release of inflammatory cytokines and inflammation of carotid artery in adventitia.

Objective To evaluate application of Steinmenn pins to assist reduction in the treatment of unstable intertrochanteric fractures with proximal femoral nail antirotation(PFNA).Methods From February 2010 to June 2013,38 unstable intertrochanteric fractures were treated by us.There were 23 men and 15 women,aged from 32 to 69 years.By Evans-Jensen classification,18 cases were type Ⅲ,13 type Ⅳ and 7 type Ⅴ.They were divided into 2 groups (n =19).Group A received reduction on a traction bed assisted by Steinmenn pins plus PFNA fixation while group B received reduction only on a traction bed plus PFNA fixation.The 2 groups were compared in terms of fracture reduction,operation time,intro-operative blood loss,fracture healing time,and Harris scores one year postoperation.Results According to the evaluation system modified by Baumgaetner et al.,the postoperative quality of fracture reduction was fine in 15 cases and fair in 4 in group A while it was fine in 9 cases,fair in 8 and poor in 2 in group B,showing a significant difference between the 2 groups (P ＜ 0.05).The operation time in group A (50.7 ± 11.9 min) was significantly shorter than in group B (63.4 ± 15.1 min),and the hip joint Harris score (89.4 ±4.4) one year after operation for group A was significantly higher than that for group B (79.6 ±6.4) (P ＜ 0.05).There were no significant differences between the 2 groups regarding intraoperative blood loss,fracture heeling time and follow-up time (P ＞ 0.05).No cases of refracture,delayed union,nonunion,or avascular necrosis of the femoral head were reported.Conclusion In the treatment of unstable intertrochanteric fractures,compared with reduction only on a traction bed plus PFNA fixation,application of Steinmenn pins to assist reduction on a traction bed plus PFNA fixation may lead to better curative efficacy due to its limited invasion,simplicity and beneficial assistance in reduction.