Objective:To optimize the water extraction and alcohol precipitation technology of Xuanfei Zhike granule .Methods:Orthogonal test was used to investigate the effects of adding water , decocting time and boiling time on the water extraction , and the effects of relative density , alcohol precipitation concentration and alcohol precipitation time on the alcohol precipitation technology by taking comprehensive score including the amount of hesperidin , the amount of tectoridin and the yield of dry cream as the indices .Re-sults:The preferred water extraction technology was as follows: added 10 times water and extracted 1.5 h firstly, and then added 8 times water and extracted twice with 0.5 h for each.The preferred alcohol precipitation technology was as follows:concentrated the wa-ter extraction to a relative density of 1.05 (measured at 60℃), slowly added 95％ethanol to 80％alcohol solution and stored 18 h at low temperature .Conclusion:The optimal water extraction and alcohol precipitation technology is stable and feasible , which can pro-vide reference for the standardized production of Xuanfei Zhike granule .

Objective To investigate the variation characteristics of peripheral platelet parameters in children with dengue fever and to assess their use and value in diagnosis,prognosis and treatment for dengue fever.Methods Platelet parameters of 69 pediatric patients with dengue fever in acute febrile phase and remission period and 103 healthy controls were compared.The difference of platelet parameters between the liver function injury group and the normal liver function group,the coagulation dysfunction group and the coagulation function normal group,the severe group and the mild group with dengue fever was compared,the causes of thrombocytopenia in children with dengue was investigated so as to better understand the situation of bone marrow producing platelets.Results P-LCR,PDW,MPV were significantly higher in patients in acute febrile phase of dengue fever,compared with the healthy subjects(P＜0.01),while PCT,PLT were significantly lower in patients in acute febrile phase of dengue fever,compared with the healthy subjects(P＜0.01).MPV was significantly lower in patients with abnormal hepatic function when compared with patients with normal hepatic function(P＜0.05),while other platelet parameters did not show significant differences(P＞0.05).PLT and PCT were significantly lower in the severe patients than those in the mild patients(P＜0.05),while other platelet parameters did not show significant differences(P＞0.05).Peripheral platelet parameters did not show significant differences between patients with or without abnormal coagulation function (P＞0.05).Conclusion Peripheral platelet parameters may play a role in the diagnosis,treatment and prognosis of dengue fever in children.

Objective To investigate the dynamics of white blood cell count (WBC),prealbumin(PA),high sensitive C-reactive protein (hs-CRP),and vascular endothelial growth factor (VEGF) in hand-foot-and-mouth disease (HFMD) complicated with or without encephalitis and in vitro inhibitory effects of artesunate on VEGF secretion of mononuclear cells from HFMD patients.Methods Peripheral blood mononuclear cells from healthy control group,HFMD group and HFMD combined with encephalitis group were isolated by Ficoll density gradient centrifugation and treated with different concentrations of artesunate (25,50,100 mg/L).The expression of VEGF in the supernatant was examined by ELISA double antibody sandwich method.The levels of WBC,PA and hs-CRP in the three groups were also detected.Multiple samples were compared by one-way analysis of variance.Multiple comparison was performed by Dunnett T3 test.Correlation of two variables was analyzed by Spearman correlation test.Results There were significant differences in the levels of WBC,hs-CRP,PA and VEGF between the HFMD combined with or without encephalitis group and health control group (F=172.69,366.02,166.32 and 5 941.89,respectively,all P＜0.01).There were significant differences in the levels of VEGF secreted by mononuclear cells treated with three different concentrations of artesunate between the HFMD combined with or without encephalitis groups and health control group (F =194.265 and 4 750.69,respectively,both P＜0.01).Correlation analysis showed that the VEGF level secretion by mononuclear cells from HFMD patients with or without encephalitis were both negatively correlated with different concentrations of artesunate (r=-0.903 and-0.969,both P＜0.01).Conclusions Compared with HFMD without encephalitis,the secretion of VEGF by mononuclear cells,WBC and hs-CRP levels in HFMD complicated with encephalitis group all increase and PA decrease significantly.Artesunate can inhibit the secretion of VEGF by mononuclear cells of patients with hand-foot-and-mouth disease in a dosedependent manner in vitro.

Objective To observe the inhibition of piperlongumine in vitro on platelet aggregation and blood coagulation tests for children,to provide the experimental basis for clinical medication.Methods Venous blood samples from 30 children were randomly devided into 5 groups,and was centrifuge to separate platelet-rich plasma (PRP).After storing in 37 ℃ thermostat water bath for 5 minutes,the PRP which have been added DMSO as blank group,and added Aspirin (10 μmol/L)as control group,and added PL (20 μmol/L),PL(100 μmol/L),PL(200 μmol/L) as different concentrations of PL groups respectively,were induced by the addition of adenosine diphosphate (10 μmol/L),collagen(2.5 μg/mL) and the arachidonic acid(500 μg/mL).Then the platelet aggregation rate of the PRP from 5 groups could be measured by turbidimetry.Blood plasma isolated from venous blood was divided into 5 groups.In the PL groups,blood plasma were mixed up with PRP concentrations of which were 5,10,20 μmol/L.In the bland group,blood plasma were mixed up with DMSO (1％).In the control group,blood plasma were mixed up with heparin sodium(35 U/mL).After storing in 37 ℃ thermostat water bath for 5 minutes,fibrinogen(FIB),prothrombin time(PT),thrombin time(TT) and activated partial thromboplastin time of different groups were detected.Results Compared to the control group,the groups which were add PL with different concentrations (20,100,200 μmol/L) showed significant inhibition on platelet aggregation induced by AA and collagen(P<0.05).PL with concentrations of 100 μmol/L and 200 μmol/L showed significant inhibition on platelet aggregation induced by ADP(P<0.05).The PT,APTT,TT of blood plasma from children had been significantly prolonged by the intervention of PL 10 μmol/L and PL 20 μmol/L(P<0.05),however,no significant change of FIB was observed.Conclusion There are inhibitory effects of PL on platelet aggregation of blood plasma from children and anticoagulant activity in this study.

Objective To investigate whether single nucleotide polymorphisms(SNPs)in the ABCC4(rs7320375,rs7329490,rs7986087),FCGR2A(rs1801274)and BLK(rs2254546)region could be susceptibility locus for Kawasaki disease(KD)in children from southern China.Methods This study was performed as a case-control study.The samples,92 individuals with KD and 194 healthy controls from southern China,were collected at Guangzhou Women and Childrens′Medical Center from October,2013 to November,2015,and the SNPs were genotyped by using the Sequenom MassArray system.The genotype distribution and allele frequency of the SNPs were analyzed using chi-square test and Fisher′s exact test.Results The genotype distribution of FCGR2A(rs1801274)in patients with KD were as follows: GG 4.3％(4/92),AG 33.7％(31/92),AA 62％(57/92),correspondingly in healthy controls were GG 48.5％(94/194),AG 41.2％(80/194),AA 10.3％(20/194)respectively,and a significant difference was found between KD patients and controls(x2=98.17,P=0.000).A allele frequency of FCGR2A(rs1801274)in KD patients(78.8％,145/184)was higher than that in controls(30.9％,x2=0.120,P=0.000).The genotype distribution of BLK(rs2254546)in patients with KD were as follows: GG 67.4％(62/92),AG 28.3％(26/92),AA 4.3％(4/92),correspondingly in healthy controls were GG 52.1％(101/194),AG 43.8％(85/194),AA 4.1％(8/194)respectively,significant differences were found between KD patients and controls(x2=6.47,P=0.039).G allele frequency of BLK(rs2254546)in KD patients(81.5％,150/184)was higher than that in controls(74.0％,287/388,x2=1.553,P=0.047).Conclusions For the children in southern China,FCGR2A SNPs(rs1801274)may be associated with the susceptibility to KD,and the A allele may increase the risk of KD.BLK SNPs(rs2254546)is also found to be associated with the susceptibility to KD,and the G allele may increase the risk of KD.

Objective To study the correlation between hyperhomocystinemia and the property of cerebral artery plaque,and the pathogenic mechanism of hyperhomocystinemia resulting in acute cerebral stroke.Methods 60 patients with acute ischemic stroke were chosen.30 patients with normal homocysteine were classified as control group.The other 30 patients with hyperhomocystinemia were classified as observation group.High resolution magnetic resonance imaging(HRMRI) and multi contrast plaque imaging technique were used to measure the cerebral artery plaque burden,the fat ratio of plaques,the hemorrhage within the plaque and fibrouscap rupture.The number of inflammatory cells in the plaque and the density of the new blood vessels were identified using the transfer constant of the contrast medium.Results In the observation group,4 cases with low-risk plaques (13.33％),10 cases with middle-risk plaques (33.33 ％) and 16 cases with high-risk plaques (53.33 ％).In the control group,13 cases with low-risk plaques (43.33％),6 cases with middle-risk plaques (20.00％),11 cases with high-risk plaques (36.67％).The number of intraplaque hemorrhage were 18 cases (60.00％) in the observation group,and 8 cases (26.67％) in the control group.The number of fibrouscap rupture were 21 cases (70.00％) in the observation group,and 13 cases (43.33％) in the control group.Risk stratification of vulnerable plaques in cerebral atherosclerosis (U =-2.032,P =0.042),intraplaque hemorrhage (x2 =6.79,P =0.009) and fibrouscap rupture (x2 =4.34,P =0.037) in the observation group was higher than that in the control group.The number of inflammatory cells in plaques and the density of the new blood vessels in the observation group was (0.188 ± 0.265)Ktrans/min,which in the control group was (0.118 ± 0.183)Ktrans/min.The number of inflammatory cells in the plaque and the density of the new blood vessels in the observation group was higher than that in the control group (t =11.831,P =0.000).Conclusion Hyperhomocystinemia causes intraplaque hemorrhage,fibrouscap rupture,inflammatory cell activity and neovascularization in plaques,accelerates cerebral atherosclerosis and increases the incidence of ischemic stroke events.

Biomedical research involving human body needs to be reviewed and oversight by the Institute Review Board (IRB) is the important international rule,aimed to protect human subjects.Ethics,however,may limited the freedom and innovation of medical research.It is an important task for the medical researchers on how to make the medical research to meet the demands of ethics,to avoid the risk,and to promote the innovation for the better development of the medical research.

Objective To investigate the reference ranges of peripheral white blood cells (WBC) and five classification parame‐ters among 13 442 apparently healthy children to provide reference for clinical diagnosis .Methods A total of 13 442 apparently healthy children aged from 1 month old to 5 years undergoing the health physical examination in the healthcare department of the Guangzhou Municipal Women and Children Medical Center were chosen according to the healthy children screening standard and the exclusion standard of hematological diseases ,infection diseases and drug administration history .The WBC analysis and its parame‐ters detection were performed by using the Sysmex XS‐800i automatic blood cell analyzer .Results WBC and its five classification parameters were not accorded with the normally distribution .By the Mann‐whitney test ,no statistically significant differences in WBC ,BASO# and NEU # were detected between different genders of children (P> 0 .05) ,while the differences in LYM # , MONO# and EOS# had statistical significance(P<0 .001) ,in which the LYM # level in females was significantly higher than that in males ,while the MONO# and EOS# levels in females were lower than those in males ;By the Kruskal‐Wallis test ,WBC and five classification parameters had statistically significant differences among different age groups (P<0 .001) .The parameters of WBC ,LYM # ,MONO# and EOS# were decreased with age increase ,while NEU # was constantly increased with age increase and BASO# was decreased with age increase before 4 years old .Conclusion All factors such as gender ,age should be The inde‐pendent reference rangesl of WBC parameters among children aged from 1 month old to 7 years old should be formulated on the ba‐sis of fully considering the factors such as gender ,age .

Objective To explore the clinical significance of platelet count increasing in patients with ovarian malignant tumor . Methods 80 cases of ovarian malignant tumor patients ,80 patients with ovarian benign tumor and 80 cases of healthy women were enrolled in the study as group A ,B ,C respectively .The group A was divided into FIGOⅠ group(n=30) ,and FIGOⅡ and above group(n=50) .Platelet count was detected by using fully automatic blood cell analyzer and the serum CA125 concentrations were also measured for all the groups .Results The platelet count and CA125 concentrations in group A were higher than group B and group C(P< 0 .05) .The positive rate of platelet increasing and serum CA125 were also higher than group B and group C(P<0 .05) .There were significant differences between FIGOⅠ group and FIGOⅡ and above group in platelet count and CA125 concen‐trations .Conclusion Ovarian malignant tumor complicated with increased platelet count is common in patients of advanced stage . The increased platelet count could indicate the malignant degree of ovarian malignant tumor .

Objective To explore changes in circulating endothelial cells(CECs)numbers in neonatal hypoxemia and its clinical significance.Methods Hladovec method was used to measure the quantity of CECs in 40 cases of neonatus with different degree of hypoxemia(hypoxemia group),and 20 matched healthy newborns were enrolled as control group.Results The numbers of CECs in hypoxemia group were much more than those in control group,and the difference was statistically significant.Meanwhile,there were significant differences in CECs numbers between moderate group and mild group(t =13.43,P <0.05),as well as moderate group and severe group(t=7.698,P <0.05).Conclusion The quantity of circulating ECEs in hypoxic neonatus is correlated with the hy-poxic severity,which may be used as early diagnostic indicator for hypoxemia,and could provide new scientific evidences for early diagnosis,illness judgment and curative effect evaluation.