The Toll-like receptors are a kind of pattern recognition receptors,which are expressed in a variety of cells,playing an important role in innate immunity and subsequent acquired immunity.At the same time,they are also involved in the immunity,proliferation,apoptosis and other physiological processes of the cells.Toll-like receptors are also expressed in hematological malignancies and involved in the progress of tumor development,immune escape,resistance,which play a double-edged sword role.In this paper,basic knowledge of TLR molecule and study on hematological tumor were mainly reviewed,aiming to provide a train of thoughts for the treatment of hematologic malignancies.

Objective:To investigate the effect and mechanism of targeted B7-H3 gene silencing on the tumorigenesis and metastasis of human hematological malignancy xenograft tumor in nude mice.Methods: Real-time fluorogentic quantitative PCR (qPCR) and flow cytometry (FCM) were used to detect the expression of B7-H3 in 13 strains of malignant hematologic cells.Then,U937,Maver and Z138 cells which expressed high level of B7-H3 were screened out.Targeted B7-H3 knockdown in U937,Ma-ver and Z138 was performed by lentivirus transduction and the effect of B7-H3 silencing in stable cell lines was tested by qPCR and FCM.Injecting the nine groups subcutaneously into the nude mice to establish xenograft models after dividing the U937,Maver and Z138 into non-infected control group (CON),B7-H3 knockdown group (KD) and negative non-targeted control infected group (NC),respectively,for detecting the tumorigenicity and metastasis in vivo.Furthermore,the expression of Ki-67 in xenograft tumors was detected by immunohistochemistry (IHC).The expression of metalloproteinase 2 (MMP-2) was detected by western blot.Results: The stable B7-H3 silencing cell lines of U937,Maver and Z138 were successfully established.Compared with the NC group,the KD groups of U937,Maver and Z138 had an obviously slower tumor growth.The average tumor inhibition rates at the end of observation period were 61.83％ (F=43.78,P<0.05),59.12％ (F=36.51,P<0.05) and 67.37％ (F=40.29,P<0.05);there was no significant difference in tumor volume growth between the NC group and the CON group (P>0.05).The liver distant metastasis of all the xenograft tumor models in nude mice was the most common and the rates of distant metastasis in KD groups were significantly lower than that of the corresponding NC groups.The Ki-67 indexes of the KD groups were significantly lower than those of the relative NC groups in three cell lines (U937: 40.3％±5.2％ vs.79.1％±6.3％,q=30.31,P<0.05,Maver: 35.2％±6.4％ vs.69.6％±5.1％,q=24.82,P<0.05;Z138: 38.4％±7.1％ vs.75.7％±4.8％,q=28.07,P<0.05);there was no significant difference in the expression of Ki-67 between the NC group and the CON group (P>0.05).The expressions of MMP-2 were also significantly lower in the KD groups than in the NC groups (U937: q=14.59,P<0.05;Maver: q=9.25,P<0.05;Z138: q=11.04,P<0.05);there was no significant difference in the expression of MMP-2 between the NC group and the CON group (P>0.05).Conclusion: Targeted B7-H3 gene silencing could inhibit the tumorigenesis and metastasis of human hematological malignancy xenograft tumor in nude mice.The mechanism may be related to the down-regulation of Ki-67 and MMP-2.

Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS),primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Hart people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).

Objective To investigate the association between SLC10A1 gene mutations in c.800G>A mutation and c.356 +1098C >T mutation, and the susceptibility to HBV infection by mother-to-child transmission ( MTCT) .Methods Totally 306 individuals born to HBeAg-positive mothers with high load HBV and without receiving nucleotide analogues treatment, including 247 HBV-infected cases and 59 non-HBV-infected ones were enrolled from Southwest Hospital during May 2011 and July 2015.Blood samples were collected from all the subjects, then genomic DNA was extracted and c.800G>A mutation and c.356+1098C>T mutation of SLC10A1 were genotyped .Chi-square test (Pearsonχ2or continuity correctionχ2) was performed to identify the difference in genotypes between two groups.Results Among vaccinated individuals (55 HBV infected and 56 not infected), the frequency of genotype GA of c.800G>A mutation in non-infected ones was 14.3%(8/56), there was a tendency of increasing compared with HBV infected ones (5.5%, 3/55), but the difference was not statistically significant (χ2 =2.424, P =0.119). Similarly, the frequencies of genotypes CC, CT and TT of the c.356+1098C>T mutation in HBV infected ones were 20.0%(11/55), 47.3%(26/55) and 32.7%(18/55), while those in non-infected ones were 12.5% (7/56), 69.6% (39/56) and 17.9% (10/56), and the difference was not of statistical significance (χ2 =5.766, P=0.056).In all subjects (vaccinated and non-vaccinated), the frequency of genotype GA of c.800G>A mutation in non-HBV infected group had an increasing tendency compared with HBV-infected offspring (13.6% vs.6.9%), but the difference was not statistically significant (χ2 =2.010, P=0.156);the frequencies of genotype CC, CT and TT of c.356+1098C>T mutation in HBV infected patients were 20.2%(50/247), 49.8%(123/247) and 30.0%(74/247), while those in non-HBV-infected group were 11.9%(7/59), 69.5%(41/59) and 18.6%(11/59), and the difference was statistically significant (χ2 =7.436, P =0.024 ) .Within the HBV infected group, the frequencies of genotype GA of c.800G>A mutation were 5.5%(3/55) in vaccinated individuals and 7.3%(14/192) in non-vaccinated individuals, and the difference was not of statistical significance (χ2 =0.030, P=0.863);Similarly, the frequencies of genotype CC, CT and TT of c.356 +1098C >T mutation in vaccinated individuals were 20.0%(11/55), 47.3%(26/55) and 32.7%(18/55), while those in non-vaccinated individuals were 20.3%(39/192), 50.5%(97/192) and 29.2%(56/192), and the difference was not of statistical significance (χ2 =0.274, P=0.872).Conclusion c.356+1098C>T mutation in SLC10A1 may be associated with susceptibility to HBV infection of child born in HBeAg positive pregnant women infected with high load HBV.

The ORFK8.1 of Kaposi's sarcoma associated-herpesvirus (KSHV) was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf K8.1 was induced by isopropyl-b-D-thiogalactopyranoside (IPTG). The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis (SDS-PAGE). SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant 0RFK8.1 protein. The optimal condition of the recombinant ORFK8.1 ELISA assay was confirmed: the concentration of antigen was 5 ug/mL, the dilution of serum was 1:200. We used the ELISA method to investigate the recombinant ORF K8.1 protein's specificity, the data showed that the specificity of ORF K8.1 to detect KSHV was 100%. At the same time, 560 sera samples from Hubei province were detected by using ORFK8.1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%.

ObjectiveTo study the clinical characteristics and laboratory ifndings ofMycoplasma pneumoniae (MP) and EBV infection in children and provide reference for clinical diagnosis and treatment.MethodsOne hundred and twenty two (122) hospitalized children with pathogen detection of MP and EBV double positive in hospitalized children with pneumonia from May 2013 to April 2014 (n=2213) were recruited as mixed infection group. In the mixed infection group, patients were further devided into high EBV mixed infection group if the EBV DNA copies were more than 1.0×104 copies/ml and the low EBV mixed infec-tion group when EBV DNA copies were less than 1.0×104 copies/ml. And another 45 hospitalized children with MP pneumonia were rectuited as control group. Clinical data and laboratory ifndings of all children were collected and analyzed.Results The mixed infection rate of MP and EBV was 5.51% (122/2213). As children getting older, the incidence of mixed infection was increased (χ2=84.08,P<0.001). And the mixed infection incidence in the lobar pneumonia group was signiifcantly higher than the bronchopneumonia group (χ2=37.44,P<0.001). The incidence of ALT and CK-MB elevated, lobar pneumonia, average fever days and hospitalization days in mixed infection with the high EBV copies group were signiifcantly higher than those in the low EBV copies group and the control group (bothP<0.05). The incidence of ALT and CK-MB elevated, average fever days and hos-pitalization days in mixed infection with the high EBV copies group were signiifcantly higher than those of the low EBV copies group and the control group (allP<0.05).ConclusionThe mixed infection of MP and EBV could aggravate the injury both in and out of the lung. Number of EBV copies plays an important role in the degree of injury both inside and outside the lung due to pneumonia with mixed infection of MP and EBV. When a patient with MP pneumonia complains with severe clinical symptoms and obvious injury outside the lung, EBV detection, especially quantitative detection of EBV DNA copies could be beneifcial for clinical diagnosis and treatment.

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.

Objective To analyze the clinical features of infants infected by respiratory syncytial virus (RSV) or human rhinovirus (HRV) in lower respiratory tract in Suzhou area based on the month age and the month of the year.Methods From January 2013 to December 2015,2 206 nasopharyngeal aspirates specimens were collected from the infants with lower respiratory tract infection.Direct immunofluorescence assay was performed to test RSV.Reverse transcription-polymerase chain reaction(RT-PCR) method was used to test HRV.The medical history was collected and pulmonary function tests were performed in some infants who were infected with RSV and HRV.Results In 2 206 cases,total RSV positive rate was 19.90％ (439/2 206 cases) and simple RSV infection positive was detected in 399 cases.Total HRV positive rate was 14.14％ (312/2 206 cases),in which simple HRV infection positive was detected in 250 cases and the detection rate of RSV was significantly higher than that of HRV(x2 =25.88,P ＜0.05).The incidence rate of wheezing in simple RSV infection was 68.17％ (272/399 cases),which was significantly higher than that of simple HRV infection (42.80％,107/250 cases) (x2 =11.174,P ＜ 0.05).RSV infection was frequent from November to February of the next year in which the detection rate in December was highest with the proportion of 50.00％ (99/198 cases) while the rate in June was only 0.57％ (1/175 cases).The detection rate of HRV was 22.86％ (40/175 cases),20.47％ (35/171 cases) and 20.33％ (25/123 cases) in June,July and September respectively.The detection rate of HRV was lower during December to February of the next year.In January,the detection rate was only 4.68％ (11/235 cases),which was the lowest in the whole year.The detection rates of RSV were 33.33％ (4/12 cases),25.21％ (118/468 cases),23.46％ (84/358 cases) and 23.81％ (60/252 cases) in the age group of 28 d-1 month,＞ 1-2 month,＞ 2-3 month and ＞ 3-4 months respectively.Up to the age of 4 months old,the detection rate decreased gradually,and with the increase of age and the detection rate in ＞ 7-8 month group was only 10.96％ (16/146 cases).The detection rate of HRV was 0 (0/12)and 9.40％ (44/468 cases) in the age group of 28 d1 month,＞ 1-2 month,respectively.After 2 months age old,the detection rate fluctuation ranged from 13.22％ to 16.67％.The incidence rate of severe RSV infection was 12.30％ (54/439 cases) and the incidence rate of severe HRV infection was 5.13％ (16/312 cases).Increased respiratory rate was more common in patients with severe RSV infection while severe HRV infection in infants were accompanied by multiple lobar involvement.After RSV infection,the incidence rate of pulmonary function damage was 89.03％ (276/310 cases).After HRV infection,89.27％ (183/205 cases)of the infants suffered from pulmonary function damage.Both RSV and HRV infection might cause pulmonary function damage.Conclusions RSV and HRV are the major pathogens in infants of Suzhou areas.The incidence of RSV-induced wheezing is significantly higher than that of HRV.RSV is detected positive mainly in winter and early spring and the infants within 4-month old are susceptible population.HRV is detected positive mainly in June,July and September and the infants older than 2 months are susceptible population.The incidence of severe RSV infection is significantly higher than that of HRV.Severe RSV infection may cause increased respiratory rate and severe HRV infection mainly cause multiple lobar involvement.RSV and HRV infection may cause pulmonary function damage.

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 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.