Objective:To explore the improvement of the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection and conduct safety tests including abnormal toxicity test,allergic reaction test,hemolysis and coagulation test.Methods:Ginkgo Leaf Extract and Dipyridamole Injection from 3 different manufactures(A,B and C)were tested respectively through abnormal toxicity test and acute toxicity test in mice,active systemic anaphylaxis test in guinea pigs and hemolysis test in vitro.Five mice were used in each batch for abnormal toxicity test according to the abnormal toxicity test method in general notice of the Chinese Pharmacopoeia 2020 Volume Ⅳ(1141),and 50 mice were selected in each batch for acute toxicity test to determine the median lethal dose(LD50)or maximum tol-erable dose(MTD)of Ginkgo Leaf Extract and Dipyridamole Injection,which were used to establish the method of abnormal toxicity experiment.The anaphylaxis of Ginkgo Leaf Extract and Dipyridamole Injection was evaluated by active systemic anaphylaxis test in guinea pigs,which was used to establish the method of allergic test.The hemoly-sis test of Ginkgo Leaf Extract and Dipyridamole Injection was studied by conventional tube method in vitro(macro-scopic observation)and improved hemolysis method in vitro(spectrophotometric method),which were used to establish the method of hemolysis and coagulation test.Results:① In manufacture A,the results of abnormal toxicity test were showed that LD50 was20.8 mL·kg-1and MTD was 16.5 mL·kg-1.No death or abnormal reac-tions were observed in mice tested for abnormal toxicity of 2 manufactures(B and C),and MTD was 50 and 40 mL·kg-1,respectively.②The no-observed-adverse-effect dose of Ginkgo Leaf Extract and Dipyridamole Injec-tion from 3 manufactures to guinea pig intravenous was 0.83 mL·kg-1,and no allergic reaction symptoms were observed when Ginkgo Leaf Extract and Dipyridamole Injection was diluted 4 times to challenge the sensitized guinea pigs(equivalent to human clinical dosage).③Differences were observed in the hemolytic effects of Ginkgo Leaf Extract and Dipyridamole Injection from 3 manufactures,but no obvious hemolytic reaction occurred when it was diluted 1.2 times(equivalent to 5%of the maximum clinical concentration).Conclusion:It is recommended to add abnormal toxicity test,allergic reaction test,hemolysis and coagulation test in the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection as safety test items to control the risk.The proposed method is diluting Ginkgo Leaf Extract and Dipyridamole Injection by 5 times,4 times and 1.2 times to perform abnormal toxicity test,allergic reaction test,hemolysis test and coagulation test respectively.

Objective:To explore the improvement of the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection and conduct safety tests including abnormal toxicity test,allergic reaction test,hemolysis and coagulation test.Methods:Ginkgo Leaf Extract and Dipyridamole Injection from 3 different manufactures(A,B and C)were tested respectively through abnormal toxicity test and acute toxicity test in mice,active systemic anaphylaxis test in guinea pigs and hemolysis test in vitro.Five mice were used in each batch for abnormal toxicity test according to the abnormal toxicity test method in general notice of the Chinese Pharmacopoeia 2020 Volume Ⅳ(1141),and 50 mice were selected in each batch for acute toxicity test to determine the median lethal dose(LD50)or maximum tol-erable dose(MTD)of Ginkgo Leaf Extract and Dipyridamole Injection,which were used to establish the method of abnormal toxicity experiment.The anaphylaxis of Ginkgo Leaf Extract and Dipyridamole Injection was evaluated by active systemic anaphylaxis test in guinea pigs,which was used to establish the method of allergic test.The hemoly-sis test of Ginkgo Leaf Extract and Dipyridamole Injection was studied by conventional tube method in vitro(macro-scopic observation)and improved hemolysis method in vitro(spectrophotometric method),which were used to establish the method of hemolysis and coagulation test.Results:① In manufacture A,the results of abnormal toxicity test were showed that LD50 was20.8 mL·kg-1and MTD was 16.5 mL·kg-1.No death or abnormal reac-tions were observed in mice tested for abnormal toxicity of 2 manufactures(B and C),and MTD was 50 and 40 mL·kg-1,respectively.②The no-observed-adverse-effect dose of Ginkgo Leaf Extract and Dipyridamole Injec-tion from 3 manufactures to guinea pig intravenous was 0.83 mL·kg-1,and no allergic reaction symptoms were observed when Ginkgo Leaf Extract and Dipyridamole Injection was diluted 4 times to challenge the sensitized guinea pigs(equivalent to human clinical dosage).③Differences were observed in the hemolytic effects of Ginkgo Leaf Extract and Dipyridamole Injection from 3 manufactures,but no obvious hemolytic reaction occurred when it was diluted 1.2 times(equivalent to 5%of the maximum clinical concentration).Conclusion:It is recommended to add abnormal toxicity test,allergic reaction test,hemolysis and coagulation test in the quality standard of Ginkgo Leaf Extract and Dipyridamole Injection as safety test items to control the risk.The proposed method is diluting Ginkgo Leaf Extract and Dipyridamole Injection by 5 times,4 times and 1.2 times to perform abnormal toxicity test,allergic reaction test,hemolysis test and coagulation test respectively.

ObjectiveTo evaluate the lipid-lowering activity of Quansanqi tablets（QSQ）， an innovative new drug of Panax notoginseng. MethodMice and golden hamsters were used to establish a hyperlipidemia model by injecting egg yolk milk and feeding high-fat diets. The levels of total cholesterol （TC），triglyceride （TG），low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） were detected， and liver function indicators ［alanine aminotransferase （ALT）， aspartate amino-transferase （AST）， and alkaline phosphatase （ALP）］ of golden hamsters were detected. Hematoxylin-eosin （HE） staining was used to observe the degree of liver injury. In the experiments， a normal group， a model group， an atorvastatin calcium group， and low-， medium-， and high-dose QSQ groups （0.32， 0.64， 1.28 g·kg^-1 for mice， and 0.16， 0.32， 0.64 g·kg^-1 for golden hamsters） were set up. ResultCompared with the normal group， the acute hyperlipidemia model mice showed increased TC， TG， and LDL-C levels （P<0.01）， and the hyperlipidemia model mice showed increased TC and LDL-C levels （P<0.01）. Additionally， the hyperlipidemia model golden hamsters showed increased serum TC， TG， LDL-C， ALT， AST， and ALP levels （P<0.05， P<0.01）. HE staining indicated the presence of fat accumulation in the liver， accompanied by inflammatory reactions. Compared with the model group， QSQ of various doses could reduce TC， TG， and LDL-C levels in acute hyperlipidemia model mice （P<0.05， P<0.01）， and the high-dose QSQ could reduce TC and LDL-C levels （P<0.01） and increase HDL-C level （P<0.05） in hyperlipidemia model mice， as well as reduce TC， TG， and LDL-C levels in hyperlipidemia model golden hamsters （P<0.05， P<0.01）， especially in the first two weeks. In addition， atorvastatin calcium could further increase ALT， AST， and ALP levels （P<0.05， P<0.01） and aggravate liver function damage， while low-dose QSQ could reduce ALT， AST， and ALP （P<0.05）， and medium- and high-dose QSQ did not cause further liver function damage. ConclusionQSQ have a significant lipid-lowering effect on different hyperlipidemia model animals and can improve liver function and liver injury.

Recombinant proteins provide new means for disease treatment, while creating considerable economic benefits. Using commercial crops (mainly tobacco), cereal crops, legumes, and vegetable crops to produce recombinant proteins with medicinal value is a hot-spot for research in "molecular farming". Although many recombinant proteins have been expressed in plants, only a small number have been successfully put into use. To overcome the problems that greatly hamper the development of recombinant protein production in plants, researchers have improved expression systems to increase the yield of recombinant proteins. Starting from analyzing the problems of low yield and/or low biological activity of recombinant proteins produced by plants, the optimization strategies to solve these problems were reviewed, and future research directions for improving the yield of recombinant proteins produced by plants were proposed.
Crops, Agricultural/genetics* ; Plant Proteins/metabolism* ; Plants, Genetically Modified/genetics* ; Recombinant Proteins ; Tobacco/genetics*

Objective To evaluate the effect of second-line antiretroviral treatment (ART) on human immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS) and provide reference for subsequent HIV/AIDS treatment.Methods Two hundred and twenty-eight HIV/AIDS patients received second-line ART during January 2011 and December 2015 in Zhengzhou were included.Two hundred and forty-eight who received first-line ART during this period were randomly enrolled as control group.CD4+ T cell count and HIV RNA load before and after treatment were compared with x2 test and t test when appropriate.Results There were 228 patients (137 male and 91 female) in the second-line ART group and 248 patients (176 male and 72 female) in the control group.In second-line ART group, CD4+ T cells increased from (274±200)/μL to (476±261)/μL after an average treatment of (39.5±18.8) months.The difference was statistically significant (t=12.91, P<0.05).In control group, CD4+T cells increased from (251±199)/μL to (470±233)/μL after an average treatment of (35.7±14.7) months.The difference was statistically significant (t=14.60, P<0.05).CD4+ T cells showed no statistical difference between two groups regardless of treatment (t=1.25 and 0.26, respectively, both P>0.05).During the treatment, the rates of immunological failure were 9.6% (22/228) in second-line ART group and 12.9% (32/248) in the control group.There was no statistical difference between two groups (x2=1.251, P>0.05).Complete viral inhibition rates were 83.3% (190/228) in second-line ART group and 88.7% (220/248) in control group with no statistical difference (x2=2.881, P>0.05).Conclusions Second-line ART regimen has equivalent treatment efficacy with first-line ART.To achieve a better outcome, second-line ART regimen should be selected as an alternative option when first-line regimen fails.Compliance is the key to guarantee the success of antiviral therapy.

Objective To measure the number of lymphocytes, B lymphocytes, CD5+B lymphocytes and level of IL-10 in peripheral blood of patients with systemic lupus erythematosus (SLE), and analyze their effects in the disease. Methods In this study, 84 cases of patients with SLE were randomly selected and evaluated according to the activity index (SLEDAI). These cases were divided into low activity group (SLEDAI<9) and high activity group (SLEDAI≥9). Ten healthy individuals were selected as the control group at the same time. The number of peripheral blood lymphocytes, B lymphocytes, CD5 + B lymphocytes, erythrocyte sedimentation rate (ESR), C3, C4 and interleukin (IL)-10 levels in serum were measured respectively and the correlation between the above indexes and SLEDAI and complement levels were analyzed. Pair-wise comparison of means of groups was conducted with one-way ANOVA. Comparison between the two groups was conducted by LSD-t test. Correlations between variables were carried out using Spearman's rank correlation test. Results The total number of lymphocytes in SLE group was lower than that in normal control group ( F=7.216, P<0.001); The number of CD19+ B lymphocytes in SLE group was higher than that in normal control group (F=3.589, P=0.036). The number of CD5+B lymphocytes of peripheral blood [(2.5±0.6)%] in patients with systemic lupus erythematosus was significantly lower than that in the normal control group [(3.2 ±0.8)%], but the difference was not statistically significant (t=3.412, P=0.698). The number of CD5+B lymphocytes in the high activity group was significantly lower than that in the low activity group (t=7.365, P=0.027)and the normal control group (t=5.649, P=0.002). The number of CD5+ B lymphocytes was negatively correlated with SLEDAI score (r=-0.692, P=0.001) and positively associated with the level of complement 3 (r=0.305, P=0.038), but not with complement 4 and ESR (P>0.05). In addition, the level of serum IL-10 in whether the low activity group (t=1.935, P=0.031) or the high activity group (t=3.048, P=0.012) was all higher than the normal control group. The level of serum IL-10 in patients with systemic lupus erythematosus was positively associated with SLEDAI score (r=0.425, P=0.024) and ESR (r=0.479, P=0.008), but was negatively correlated with complement 4 (r=-0.359, P=0.031). Conclusion The total number of lymphocytes in patients with SLE decreases significantly, while B lymphocytes increases significantly. The number of CD5+ B lymphocytes and the serum IL-10 level are also changed. It maybe related to the patient's inflammatory environment, and the number of CD5+B lymphocytes and the serum IL-10 level may be associated with disease activity.

Objective To investigate the correlation between carotid artery plaque and blood pressure variability (BPV) in elderly patients with hypertension.Methods One hundred sixty elderly patients with hypertension were divided into plaque and non-plaque groups according to the results of carotid artery ultrasonography.All the patients were measured by ambulatory blood pressure monitoring.The mean blood pressure,mean pulse pressure,and blood pressure variability coefficient of two groups were calculated and compared during whole day,daytime,and nighttime.The related factors of carotid artery plaque were analyzed by multivarite logistic regression analysis.Results The 24 h systolic blood pressure standard deviation,daytime and nighttime of systolic blood pressure standard deviation,daytime diastolic blood pressure standard deviation of plaque group were higher than those of non-plaque group (P ＜ 0.05).The 24 h systolic blood pressure variation (24 h SBPV) and night systolic blood pressure variation (nSBPV) were higher than those of non-plaque group (P ＜0.01).Multivariate regression analysis results showed that carotic artery plaque was associated positively with 24 h BPV and blood pressure variability coefficient of nighttime (P ＜ 0.05).Conclusions The elderly hypertensive patients with carotid artery plaque is associated positively with 24 h systolic blood pressure variability coefficient and blood pressure variability of nighttime.

[Summary] Four patients with hyperthyroid-associated exophthalmos, myxedema, acropachy ( EMA ) syndrome, including three male patients and one female patient were diagnosed with Graves′diseases and treated by 131 I therapy. Complaints of thyrotoxicosis were presented at the onset. Tibia myxedema and acropathy appeared, and eye symptoms aggravated in two patients after anti-thyroid drug therapy and 131 I therapy. Four cases were all given clobetasol propionate, miconazole nitrate, neomycin sulfate and urea cream alone or in combination with compound betamethasone local injection treatment, and three cases were given low-dose oral prednisone treatment. Complaints of tibia myxedema and eye symptoms were significantly improved after the treatment. Therefore, we should be wary of the occurrence of hyperthyroid-associated EMA syndrome after 131 I therapy. Corticosteroid might be the effective therapy for myxedema and eye symptoms of EMA syndrome.

Objective To study diagnostic methods for early renal injury in hypertension patients. Methods Urinary microalbumin (mALB) and ? 2 microglodulin(? 2 MG) levels were measured with rate nepherometry. Total quantitative enzyme immunoassay was employed to measure urinary retinol binding protein (RBP) levels, rate for urinary N acetyl beta D glucosaminidase (NAG), and Jaffes rate for urinary creatinine (Cr). Results The levels of urinary RBP, mALB, ? 2 MG, NAG in hypertension patients were significantly higher than those in controls ( P

Objective To study CEAmRNA、CK19mRNA and CK20mRNA in peripheral blood of lung cancer patients by Real-time RT-PCR,consequently discussing those result′s clinical significance.Methods 48 lung cancer patients,8 benign lung disease patients and 30 healthy volunteers of CEAmRNA,CK19mRNA and CK20mRNA in peripheral blood were detectd by Real-time RT-PCR.Results The levels of 48 lung cancer patients CEAmRNA in peripheral blood reach (16 264?28 765) copies/ml,of which 26 patients,results are positive; The levels of 48 lung cancer patients CK19mRNA in peripheral blood reach (14 891?27 244) copies/ml,of which 27 patients,results are positive; The levels of 48 lung cancer patients CK20mRNA in peripheral blood reach (10 924?21 678) copies/ml,of which 20 patients,results are positive. Among of 8 benign lung disease patients,there is just CK20mRNA was detected in one patient,both CEAmRNA and CK19mRNA were not detected in their peripheral blood. As for those 30 healthy volunteers,there is none of the mRNA was detected. Moreover,there is a marked positive correlation between the levels of CEAmRNA,CK19mRNA and CK20mRNA,the levels of CEAmRNA,CK19mRNA and CK20mRNA were correlated to the physiological cause or degree of lung cancer.Conclusion CEAmRNA,CK19mRNA and CK20mRNA in peripheral blood can be as auxiliary index for diagnosing lung cancer micrometastasis,also can monitor the lung cancer micrometastasis by quantity it.