OBJECTIVE:To establish a method for the simultaneous determination of 7 active constituents in Tangshen qing-du granule. METHODS:HPLC was performed on the column of SHIMADZU Inert Sustain C18 with mobile phase of acetonitrile-0.1%phosphoric acid at a flow rate of 1.0 mL/min,detection wavelength was 327 nm for chlorogenic acid and caffeic acid,280 nm for baicalin,228 nm for arctiin and 276 nm for wogonoside,baicalein and wogonin,column temperature was 35℃,and injection volume was 10 μL. RESULTS:The linear range was 4.830-154.6 μg/mL for chlorogenic acid and(r=0.9998),0.750-24.1 μg/mL for caffeic acid(r=0.9997),22.859-731.5 μg/mL for baicalin(r=0.9997),8.491-271.7 μg/mL for arctiin(r=0.9993),2.471-79.0μg/mL for wogonoside(r=0.9996),6.656-213.0 μg/mL for baicalein(r=0.9994) and 2.756-88.2 μg/mL for wogonin (r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 2.0%,recoveries were 96.86%-100.82%(RSD=1.46%,n=6),98.79%-101.09%(RSD=0.93%,n=6),97.57%-101.51%(RSD=1.37%,n=6),97.76%-99.63%(RSD=0.77%,n=6),97.99%-100.12%(RSD=0.76%,n=6),96.54%-101.07%(RSD=1.87%,n=6) and 96.60%-99.59%(RSD=1.14%,n=6). CONCLUSIONS:The method is simple with good precision,stability and reproducibilty,and can be used for the simultaneous determination of 7 active constituents in Tangshen qingdu granule.

Objective To observe insulin resistance in first-degree relatives of patients with Graves disease. Methods All subjects in control group and experiment group including first-degree relatives of GD patients underwent oral glucose tolerance tests (OGTT) and insulin releasing tests then the degree of insulin resistance was analyzed. Results Blood glucose at each point of OGTT, insulin level and insulin resistance index 1 (HOMA-IR) of experiment group were higher than those in control group, while insulin activity index (IAI) and HOMA-βwere significantly lower than those in control group. Conclusion Patients insulin resistance could be found among first-degree relatives of GD patients.

Objective To study the epidemiological and clinical features of human parainfluenza virus (HPIV) infection in children in Suzhou,and to provide the evidence-based foundation for early warning,diagnosis and treatment of respiratory infection in children.Methods The sputum specimens and medical history were obtained from children with acute respiratory tract infection hospitalized at the Childen's Hospital Affiliated to Soochow University from January 2006 to December 2015.Seven kinds of common respiratory viruses including respiratory syncytial virus,influenza virus A,influenza virus B,HPIV Ⅰ,HPIV Ⅱ,HPⅣV Ⅲ and adenovirus were detected by using the direct im-munofluorescence.Mycoplasma pneumoniae(MP),chlamydia pneumoniae,human bocavirns (hBoV) were detected by using fluorescence quantitative PCR.Rhinovirus and human metapneumovirus were detected by using reverse transcription-PCR.Sputum was cultured for bacteria.Results In 21 769 cases,the detection rate of HPIV positive was 3.21％ (829 cases),among which,HPIV Ⅰ,HPIV Ⅱ,HPIV Ⅲ were respectively detected in 113 cases (0.52％),16 cases (0.07 ％) and 700 cases (3.21％),respectively.There were 378 cases of simple infection and 428 cases of mixed infection,and the mixed infection was very common in Streptococcus pneumoniae,Haemophilus influenza,MP and hBoV.There was a difference in HPIV infection among genders,and the detection rate of the boys was higher than that of girls[4.14％ (563/13 591 cases) vs.3.25％ (266/8 178 cases),x2 =11.036,P =0.001].In the 28 d-1 year old and ＞ 1-3 year old group,the detection rate of HPIV was higher[4.71％ (494/10 476 cases) and 4.21％ (244/5 793 cases),respectively].In spring and summer,there was a higher detection rate of HPIV infection.The clinical manifestations with simple infection of HPIV Ⅰ and HPIV Ⅲ were cough,fever and wheezing.The rate of fever and shortness of breath in those of HPIV Ⅰ was 71.74％ (33/46 cases),10.87％ (5/46 cases),and that in HPIV Ⅲ was 40.12％ (134/334 cases),2.10％ (7/334 cases),HPIV Ⅰ infection was more likely to cause fever and shortness of breath than those of HPIV Ⅲ,there were significant differences (x2 =16.410,P ＜ 0.001;x2 =10.177,P =0.001).Pneumonia had the highest detection rate of viral infection.Conclusions HPIV Ⅲ is the leading pathogen among the types of HPIV in the hospitalized children in Suzhou area.Among the subtypes of HPIV,the peak of HPIV infection occurs in spring and summer.The children less than 3 years old are the most susceptible to parainfluenza virus,and the HPIV detection rate is gradually declines with age.

OBJECTIVE:To optimize the water extraction technology of Yinju jiedu oral liquid,and provide reference for the industrial production of the preparation. METHODS:According to the investigation of extraction time-extraction rate curves of chlo-rohenic acid of Yinju jiedu formula and extraction rate of chlorohenic acid in Lonicera japonica and other combined medicinal mate-rials in the formula,decoction methods and time of L. japonica were determined. Using the comprehensive scores of linarin,harpa-goside,(R,S)-epigoitrin,psoralen+angelicin contents and dry extraction yield as indexes,L9(34)orthogonal test was designed to detect the effects of adding water amount,decoction time times and optimize the extraction technology of the residues and other me-dicinal materials. Verification test was conducted. RESULTS:The optimal technology was L. japonica decocted first for 30 min with 8-fold water;the residues and other medicinal materials were decocted with 8-fold water for 3 times,1 h each time;combin-ing all the syrups. In verification test,the average contents of chlorohenic acid,linarin,harpagoside,(R,S)-epigoitrin,psoralen+angelicin were respectively 34.51,10.31,1.97,0.21,9.79 mg/g(RSD=1.24％,1.19％,1.40％,1.71％,1.28％,n=3);aver-age dry extraction yield was 25.4％(RSD=1.64％,n=3);average extraction rate of chlorohenic acid was 78.95％(RSD=1.24％,n=3). CONCLUSIONS:In the optimized water extraction technology,both the extraction rate of chlorohenic acid and contents of other ingredients are relatively high. The technology is stable and feasible.

Objective To study the characteristics of etiology of lobar pneumonia in hospitalized children.Methods Medical history and sputum specimens were collected from 1 179 hospitalized children with lobar pneumonia from January 2006 to December 2015.Multiple pathogenic joint detection combined with the history data were used for analysis.Seven kinds of common respiratory virus were detected by direct immunofluorescence.Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP) and human Bocavirus (hBoV) were detected by fluorescence quantitative polymerase chain reaction (PCR).Human Rhinovirus (HRV) and human Metapneumovirus (hMPV) were detected by reverse transcription PCR.Aspirates were cultured for bacteria.MP specific antibody IgG and IgM were tested by enzyme-linked immunosorbent assay (ELISA).Positive rates of each group were compared by χ2 test or Fisher exact test.Results Total etiology detection rate of lobar pneumonia in hospitalized children was 83.9% (989/1 179).The etiology detection rate of MP, virus, bacteria and streptococcus pneumoniae (SP) were 74.0%, 14.2%, 18.3% and 12.2%, respectively.The virus detection rate in 1-3 years old group was the highest, and that in ≥6 years old group was lower than other group (χ2=70.095, P<0.01).The MP detection rate increased with age (χ2=119.777, P<0.01).The bacteria detection rate in ≥6 years old group was significantly lower than those of <1 years old group, 1-3 years old group and 3-6 years old group (χ2=8.939, 14.319, 45.738, all P<0.01).The detection rates of total virus, MP, bacteria and mixed infection had no statistical difference in the four seasons (all P>0.05).The MP detection rate was above 70% in every season.The detection rates of SP and hBoV were basically the same in every season.The detection rate of HI was higher in spring, Pinf 3 and SA were higher in summer, HRV was higher in autumn, and respiratory syncytial virus (RSV) and moraxella catarrhalis (MC) were higher in winter.Conclusions Lobar pneumonia occurs more common in elder children.MP is the major pathogen of lobar pneumonia, and SP is the second.The MP detection rate increases with age.The pathogen detection rate varies with age, but the effect of seasonal factor is not obvious on pathogen detection in lobar pneumonia.

Objective To analyze the correlation between response to hepatitis B virus (HBV) vaccine and cellular immunity and clinical characteristics in children with respiratory infection.Methods Nine hundred and sixty children in Department of Respiratory in Children's Hospital of of Soochow University,who were over 7 months old and had full course of HBV vaccination between January and December 2015 were enrolled in this study.Peripheral blood (1-2 mL) was collected,and antigen-antibody of HBV was detected by using enzyme-linked immunosorbent assay and PCR included HBV surface antigen,hepatitis B antibody,HBV e antigen,HBV e antibody,HBV core antibody,and HBV nucleic acid.According to the results,these children were divided into 4 groups:non response group,low response group,normal response group and high response group according to their responses to HBV vaccine.Cellular immunity was detected by using flow cytometry and patients' clinical data was collected.Results There was no statistical differences of CD3 + CD4 +,which were (3.43 ± 0.28) ％,(3.42 ± 0.30) ％,(3.43 ± 0.36) ％ and (3.52 ± 0.29) ％,among the four groups (F =0.520,P =0.669).CD3 + CD8 + in non response group was (3.18 ±0.28)％,which was significantly higher than that in low response group,normal response group and high response group [(3.08 ± 0.36)％,(3.05 ±0.34)％,(2.93 ±0.30)％],the differences were significant (all P＜0.05);CD4/CD8 in non response group (0.26 ± 0.43) were significantly lower than that in normal response group (0.40 ± 0.50),the differences were significant (P =0.001).There was no significant difference of CD3 +,CD3 + CD8 + and CD4/CD8 among low response group,normal response group and high response group (all P ＞ 0.05).CD3-CD19 + and CD19 + CD23 + level were lowest in non response group [(3.00 ± 0.57) ％,(2.25 ± 0.67) ％] and highest in high response group [(3.33 ± 0.45) ％,(2.57 ± 0.38) ％],the differences were significant (all P ＜ 0.05).Among the 4 groups,children in normal response group had the shortest average hospitalization days [(1.88 ±-0.31) d],which was significantly shorter than that in non response group,low response group and high response group [(1.96 ± 0.39) d,(1.95 ± 0.38) d,(1.96 ±0.15) d],the differences were significant (all P ＜0.05),there was no significantly difference of average hospitalization days among other 3 groups (all P ＞ 0.05).Proportion of severe pneumonia was significantly higher in non response group [6.1％ (22/363cases)] and high response group [13.3％ (2/15 cases)] compared to those in normal response group [2.6％ (7/274cases)],the differences were statistically significant (x2 =4.417,P =0.036;x2 =5.476,P =0.019).The total white blood cell number was lowest in non response group (F =4.695,P =0.003).Platelet number was increased with higher degree of response to HBV (F =6.598,P ＜ 0.001).Conclusions Cellular immunity is lower in respiratory infection children with non response or low response to HBV vaccine.After they have respiratory infection,children with non response to HBV vaccine may have a longer course of disease and worse condition.

Objective To explore the non-bacteria pathogens of acute laryngitis in children. Methods The clinical data and sputum sample were collected from 325 patients hospitalized due to acute laryngitis in consecutive 10 years from January 2006 to December 2015 . The multiple non-bacteria pathogens were detected and analyzed with clinical data. Seven types of respiratory viruses were detected by direct immunolfuorescence. Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Boca virus (HBoV) were detected by lfuorescence quantitative PCR. The rhinovirus (HRV) and human metapneumovirus (hMPV) were detected by RT-PCR. Venous blood was collected within 24 h after hospitalization and 7-10 d after treatment. The MP antibody of IgG and IgM were detected by ELISA. Results The detection rate of non-bacteria pathogens was 46 . 2%in 325 children with acute laryngitis ( 150/325 ), including 76 cases ( 23 . 4%) of virus and 99 cases ( 30 . 5%) of MP. Virus detection rate in 1-3 year old children was obviously higher than in 0-1 year old children and over 3 years old children (χ2?=?9 . 527 , P=?0 . 009 ). With the increase of age, the detection rate of MP increased gradually (χ2?=?10 . 132 , P=?0 . 006 ). The detection rates of RSV and hBoV were higher in under 3-year-old children. The detection rates of virus in winter and spring were signiifcantly higher than those in summer and autumn (χ2?=?5.064, P=?0.024). The detection rates of MP in winter, spring, summer, and autumn was 13.1%, 25 . 0%, 38 . 2%, and 44 . 9%respectively, and the MP detection rates were increased gradually over seasons (χ2?=?4 . 438 , P=?0 . 035 ). The detection rate of RSV was higher in winter, and hBoV was higher in summer. Conclusion Acute laryngitis mainly occurred in children under 3-years-old children, and the detected non-bacteria pathogens were different among different ages and seasons. Virus was the major pathogens in young children, while MP was more common in older children.

Objective To explore the level of expression, clinical signiifcance of sB 7-H 3 in the bronchoalveolar lavage lfuid (BALF) of refractory Mycoplasma pneumoniae (MP) pneumonia (RMPP) in children and the relationship between sB7-H3 and various cytokines. Methods The BALF of forty-three hospitalized children with RMPP (RMPP group) were collected for the diagnosis and treatment. Thirteen cases were lavaged only once and the other thirty cases had collected the BALF twice. The BALF of iffteen hospitalized children with bronchial foreign body were collected as control group. The expression levels of sB 7-H 3 , IL-1β, IL-2 and IL-36 in the BALF were detected by enzyme-linked immunosorbent assay. The expression levels of sB 7-H 3 , IL-1β, IL-2 and IL-36 in the BALF at the acute phase were compared with control group and the group after treatment. Analyzed the correlation between the level of sB 7-H 3 and IL-1β, IL-2 , IL-36 in the BALF of RMPP children at acute stage. Results The levels of sB 7-H 3 , IL-1β and IL-36 in the BALF of the ifrst lavage group were higher than those of single lavage group and control group (all P<0 . 05 ). The levels of sB 7-H 3 , IL-1β, IL-2 and IL-36 in the BALF of single lavage group were higher than those of control group (all P<0 . 05 ). The levels of sB 7-H 3 , IL-1β, IL-2 and IL-36 in the BALF of the second lavage group were lower than those of the ifrst lavage group (all P<0 . 05 ).The levels of sB 7-H 3 , IL-2 in the BALF of the second lavage group were higher than those in the control group (both P<0 . 05 ), but the levels of IL-1β, IL-36 in the BALF showed no difference between the second lavage group and the control group (both P>0 . 05 ). The levels of sB 7-H 3 had positive correlation with the levels of IL-1β, IL-2 and IL-36 (all P<0 . 001 ). Conclusions sB 7-H 3 may control the secretion of IL-1β, IL-2 and IL-36 , and participate in immune response and lung injury after MP infection, which may lead to occurrence and development of RMPP.

Objective To analyze the epidemiological characteristics of Mycoplasma pneumoniae (MP) infection in children with respiratory tract diseases ,and to provide scientific basis for clinical diagnosis and treatment and to formulate control measurements for the administrative department of public health .Methods Sputum specimens of 20 021 cases of hospitalized children with respiratory tract diseases from October 2005 to December 2014 in Suzhou were collected .MP DNA was detected by fluorescence quantitative polymerase chain reaction .At the same time ,venous blood was collected within 24 h after admission and 7-10 d of treatment .Specified MP antibodies IgG and IgM were tested by enzyme-linked immunosorbent assay to analyze the detection rate of MP . The positive rates between groups were compared using chi-square test or Fisher exact test .Measurement data were compared using Wilcoxon test .Results The MP infection rate was 36 .08% (7 224/20 021 cases) in 20 021 children .The MP infection rate of girls was 40 .81% (3 057/7 490) ,which was significantly higher than that of boys (33 .25% [4 167/12 531] ,χ2=116 .20 ,P<0 .01) .The MP infection rates of children at the age of less than six months ,6 months to 1 year old ,1-3 years old ,3-7 years old and older than 7 years old were 18 .35% ,29 .39% ,43 .93% ,54 .10% and 64 .48% ,respectively ,which increased with age (χ2 =1 949 .65 , P<0 .01) .The MP infection rates in spring ,summer ,autumn and winter were 31 .97% ,41 .57% , 40 .88% and 29 .90% , respectively . The MP infection rate of children in summer and autumn was significantly higher than that in spring and winter (χ2 =234 .61 , P<0 .01) .The MP infection rate was highest in the autumn of year 2008 (55 .07% ) and lowest in the spring of year 2010 (18 .48% ) for the decade .The MP infection rate showed fluctuations with different degrees in four seasons except in 2007 . In the past ten years ,the MP infection rate in Suzhou area was at a higher level in 2008 ,2009 ,2012 and 2013 ,which were 46 .03% ,46 .60% ,39 .28% and 47 .40% ,respectively .The MP infection rate was the lowest (25 .24% ) in 2011 in the decade ,and maintained around 30% in the rest years .Conclusions The MP infection rate in children with respiratory tract diseases is at a high level in Suzhou area .The MP infection rate of girls is higher than that of boys .MP infection could occur among all age groups ,and the MP infection rate increases with age .MP infection rate peaks in summer and autumn .MP infection has a small prevalence every two or three years ,which could sustain about two years .

Objective To study the pathogenic etiology between nasopharyngeal aspirates (NPA) and bronchoalveolar lavage lfuid (BALF) in children with lower respiratory infection. Methods Multiple pathogen in NPA and BALF from 210 cases with lower respiratory tract infection was detected. Seven common respiratory virus (respiratory syncytial virus, adenovirus, in-lfuenza virus A, inlfuenza virus B, parainlfuenza 1, parainlfuenza 2, parainlfuenza 3) were detected by direct immunolfuorescence assay. MP, CP and HBoV were detected by lfuorescence quantitative PCR.HRV and hMPV were detected by RT-PCR. Aspirates were cultured for bacteria. The results of pathogen detection in secretions of upper and lower respiratory tract were analyzed. Results Total positive detection rate of NPA and BALF in 210 cases was 91.9%(193/210), which is higher than that in NPA 75.2%(158/210) and that in BALF 85.2%(179/210). Bacteria detection rate in NPA was 13.3%(28/210), and 8.6%(18/210) in BALF, without signiifcant difference (P=0.118). Bacteria detection rate in NPA and BALF was of poor consistency (Kappa=0.262). Virus detection rate in NPA was 24.3%, which is higher than that in BALF15.2%. BALF-MP detection rate was 77.6%(163/210), signiifcantly higher than that in NPA 53.3%(112/210). There are 95.5%(107/112) cases with positive results in NPA-MP detec-tioncan also be detected in the BALF-MP. MP copies in BALF were signiifcantly higher than that in NPA (4.28×106 vs. 1.31×105), and its positive rate in NPA was still higher than that in BALF. MP detection rate in NPA in children with clinical course of longer than two weeks was much lower than those with clinical course of two weeks or less. Conclusions The pathogen detection of virus and MP in NPA can be used as a reference for lower respiratory tract infection. The joint detection of NPA and BALF can improve the detection power. The sensitivity of virus detection in NPA is higher than that in BALF. NPA pathogen detection of virus and MP is of great important evidence-based medicine in the diagnosis of lower respiratory infection. MP detection rate and its copies in BALF are signiifcantly higher than that in NPA. BALF detection is the supplement of pathogen diagnosis in severe or refractory lower respiratory infections.