Clinical data of a case of Munchausen syndrome by proxy (MSBP) admitted to the Department of Pediatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology in November 2020 were retrospectively analyzed.The 4 years and 4 months old female patient presented with retrosternal and abdominal pain for 1 month, and aggravated with multi-organ pain for 20 days.She complained about the retrosternal pain with acid reflux, pain in the teeth, esophagus, and abdomen, etc.During the hospitalization, she frequently complained of multi-organ pain.Her mother repeatedly declared her painful hip joint and she often cried for pain at night, and even could not walk.However, the clinical examination showed no obvious abnormalities.Combining characteristics of the patient and her caregiver, the patient was confirmed as MSBP.It is suggested that MSBP in children should be concerned in cases with complicated severe chief complaints, frequent medical visits, and a strong willing to see a doctor or be hospitalized by their caregivers, but normal physical and auxiliary examination findings.

Activated phosphoinositide 3-kinase-delta syndrome(APDS) is a rare autosomal dominant primary immunodeficiency disease.According to mutation types, APDS is divided into two types, APDS1 and APDS2.APDS1 patients have more susceptibility to develop bronchiectasis, sinusitis, hepatomegaly, splenomegaly, asthma, autoimmune or inflammatory diseases, and are more frequently infected with Streptococcus pneumoniae and Haemophilus influenzae, while APDS2 patients are more prone to get pneumonia, eye infection, and lymphadenopathy, malignancy, neurological and growth retardation.Among the immunological features, the T cell count of APDS1 is significantly low, and APDS2 is more obvious to appear elevated IgM levels.Rapamycin is beneficial for both types of APDS, and Leniolisib is better tolerated in patients with APDS1.This article reviews the differences in pathogenesis, clinical manifestations, immunological characteristics, and treatment between APDS1 and APDS2 to improve the understanding by clinicians.

Innate immunity is on the frontline of fight against pathogenic microorganism invasion .As a DNA sensor, absent in melanoma 2 (AIM2) is an important member of innate immune system.It can recognize dsDNA of pathogenic microbes to form AIM 2 inflammasomes , which facilitates defending and clearing the invasion of pathogens by activating caspase-1 dependent pyroptosis and the mature of IL-18 and IL-1β.AIM2 inflammasomes play an important part in responding to Listeria monocytogenes, Francisella tularensis, Streptococcus pneumoniae, Mycobacterium tuberculosis, Aspergillus fumigatus, vaccinia virus , murine cytomegalovirus , and hepatitis B virus infections .This paper introduces the components of AIM 2 inflammasomes and summarizes its function in defending the invasion of pathogenic microorganisms .

Objectives To investigate the effects of lipoxin receptor agonist BML-111 on IFN-βand IE86 mRNA expression of macrophages infected by human cytomegalovirus (HCMV). Methods Macrophages were infected with HCMV (MOI=0.5), and the cultured cells were randomly divided into control group, HCMV group, HCMV+BML-111 group, and HCMV+MP group. The cells were collected at 0,1,2,4,8 and 12 h after infection, and the levels of IFN-βand IE86 mRNA were tested by real-time PCR. Results Compared with HCMV group, the levels of IFN-βmRNA in HCMV+BML-111 group increased significantly (P < 0.05), while the levels of IFN-βmRNA in HCMV+MP group decreased significantly (P < 0.05); Compared with HCMV group, there were no significant differences of the levels of IE86 mRNA in HCMV+BML-111 group (P>0.05), while the levels of IE86 mRNA in HCMV+MP group increased significantly (P < 0.05). Conclusion BML-111 exerts antiviral activity by promoting the expression of IFN-βmRNA at the early stage of HCMV infection.

BACKGROUND:The anti-inflammation and protective effects of lipoxin have been verified in several immunity-related disease models. Preliminary studies of our research group have shown that, lipoxin receptor agonist BML-111 has negative regulation effects on the human cytomegalovirus (HCMV)-induced immunological injury. However, the effect of BML-111 on the HCMV replication remains unclear.
OBJECTIVE:To observe the influence of lipoxin receptor agonist BML-111 on HCMV replication and proliferation in THP-1 macrophages and human embryonic lung fibroblasts.
METHODS:THP-1 macrophages were infected by HCMV AD169 strain, and were divided into three groups:mock infection, HCMV infection, HCMV+BML-111. The final concentration of BML-111 was 100 nmol/L. cells in each group were col ected at 0, 1, 2, 4, 12, 36, 48 hours, the mRNA levels of IE86 and pp65 in the THP-1 macrophages were tested by RT-PCR method. Human embryonic lung fibroblasts were infected with HCMV (MOI=0.1), and were divided into two groups:HCMV infection and HCMV+BML-111. The patho-morphous changes of human embryonic lung fibroblasts were observed under light microscope, and the cellnumber was measured. The infective virus titer changes in human embryonic lung fibroblasts were examined by plaque assay.
RESULTS AND CONCLUSION:After the macrophages were infected by HCMV, compared with the mock infection group, the mRNA levels of IE86 and pp65 in the HCMV group and HCMV+BML-111 group were increased significantly;compared with the HCMV infection group, the mRNA levels of IE86 and pp65 in the HCMV+BML-111 group were increased significantly in the early stage (within 4 hours) after infection, but the pp65 mRNA levels were decreased significantly in the medium and late stages (24-72 hours) after infection. After human embryonic lung fibroblasts were infected by HCMV, the degree of the patho-morphous in the HCMV+BML-111 group reached 100%2 days earlier than the of HCMV infection group. The infective virus titer reached the peak 2 days earlier than the HCMV infection group, but no significant difference was found between the two groups. BML-111 accelerates the replication of HCMV in the early stage of infection, but inhibits the expression of pp65 gene in the late stage. BML-111 has no impact on the proliferation of the infective HCMV titer in vitro.

Objective To explore the correlation between the expression of IL-17A and the degree of spleen damage in acute mouse cytomegalovirus(MCMV) disseminated infection in vivo and to understand the mechanism about how IL-17A involved in the pathological damage of the spleen in MCMV infection.Methods An acute disseminated MCMV infection model was established in mice.BALB/c mice were randomly divided into two groups.Mice in group one were infected with MCMV Smith to establish disseminated infection.Mice in another group were sham-infected control.Three mice from each group were randomly chosen to be sacrificed on days 3,7,14 and 28 after the infection.Viral titers in spleen tissues were determined using a standard plaque assay.The expression of IL-17A mRNA and MCMV mRNA in the splenocytes were measured by RT-PCR.The expression of IL-17A in spleen tissues was observed by immunohistochemical staining.The pathology of the infected mice was assessed by histological examination of H&E stained spleen sections.Results Viral titers and MCMV mRNA in the spleen peaked on day 3,but quickly diminished on day 7.Virus was no longer detectable in the spleen on day 14 after the infection.The expression of IL-17A mRNA was significantly increased during the acute infection and reached the highest level on day 14,then decreased on day 28.It is significantly higher than that of the mock infection group.Immunohistochemistry assay also indicated that the expression of IL-17A in spleen tissue gradually increased to climax on day 14,then decreased on day 28.Accordingly,the pathological damages of spleen tissue in the infected mice deteriorated until day 14,then showed signs of recovery on day 28.The most severe pathological injury of spleen tissue and the highest expression of IL-17A appeared in the same period of time.Conclusion Our results showed a close correlation between IL-17A and the pathological damage in spleen.Thus,IL-17A may contribute to the spleen pathological damage during the acute disseminated MCMV infection.

ObjectiveTo observe the changes of AIM2 ( absent in melanoma 2) inflammasome during early murine cytomegalovirus (MCMV) infection.MethodsBALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminated infection,the other was sham-inoculated control.On days 1,3,5 and 7 of the experiment,three mice of each group were randomly chosen to be killed separately.The expression of AIM2,ASC and caspase-1 in splenic macrophages was detected by Western blot,the levels of IL-1β and IL-18 in sera were measured by double antibody sandwich ELISA,and the viral titers in salivary gland tissues were quantified by a standard plaque assay.Results The MCMV titers in salivary gland tissues were gradually increased in MCMV-infected mice on days 3,5 and 7,while the expressions of AIM2 in macrophages were began to increase on day 1 and significantly increased and reached the highest level on day 3 but gradually decreased afterwards.The relative intensity of AIM2 on day 3 differed significantly between the MCMV-infected mice and the controls (1.121±0.243 vs 0.240±0.046,P＜0.01,t test),as did ASC ( 1.318±0.333 vs 0.248±0.090,P＜0.01 ) and caspase-1 ( 1.085±0.243 vs 0.247±0.064,P＜0.01 ).Meanwhile,the levels of IL-1β and IL-18 in MCMV-infected mice were (112.72±5.20) pg/ml and (42.74±4.23) pg/ml,and the levels were significantly higher (P＜0.01 ) than those in controls [ (47.86±4.35) pg/ml and (22.60±2.82) pg/ml].ConclusionThese results demonstrate that AIM2 inflammasome is activated in macrophages during early MCMV infection and could be as a therapeutic target for CMV-induced diseases.

Objective To investigate the nature of Th17 cells in murine cytomegalovirus(MCMV)infection during the acute stage,we characterized the frequency of IL-17A-producing CD4 T cells and the level of Th17 cytokines,IL-17A,in MCMV-infected mice.Methods BALB/c mice were randomly divided into two groups.One was infected with MCMV Smith for establishing disseminative infection; the other was sham-inoculated control.On day 3,7,14 and 28 of the experiment,three mice of each group were randomly chosen to be killed separately.Real-time PCR was used to detect MCMV loads in organs of MCMV-infected mice,the pathology of spleen was observed by HE staining.The frequency of CD4+IL-17A+ T cells in total splenocytes of mice was detected by flow cytometry.The level of IL-17A in culture supernatants of splenocytes was measured by double antibody sandwich ELISA.Results MCMV loads in salivary gland reached the peak on day 14 after MCMV infection,the most severe spleen injury was also shown on day 14,the frequencies of CD4+IL-17A+ T cells in total splenocytes increased significantly( all P＜0.01 ) in MCMV-infected mice than those in controls,and reached the peak on day 14 ( 1.14％ ±0.09％ vs 0.19％ ±0.04％,t =17.551,P=0.000).The levels of MCMV-specific IL-17A in culture supernatants were increased dramatically in MCMV-infected mice than those in controls on day 14 [ (81.98± 12.37) pg/ml vs (44.96±8.44)pg/ml,t=4.281,P=0.006].In MCMV-infected mice,correlation was positive between the levels of MCMV-specific IL-17A in culture supernatants and MCMV loads in salivary gland tissues (r=0.54,P＜0.05 ),the levels of IL-17 A in culture supernatants were higher in more severe spleen injury.Conclusion Thl7 cells and IL-17A were involved in the immunity response during acute MCMV infection.They may correlate with the persistence of MCMV and the pathology of spleen in infected mice.

Objective To establish the murine congenital infection model by MCMV and observe the pathological changes and infection status of brain tissue.Methods After anesthesia,mice who were pregnant 11-13.5 days (E11-13.5 d) were intra-amniotic injected one uterus by one with virus (MCMV K181 suspension,1 μl,1×103 PFU).The control group of the same period was intra-anmiotic injected with culture medium DMEM (1 μl).Carefully reset the uteruses and close the abdomen.After 5 days of separated feeding,kill the pregnant mice,take the fetus out of the uterus,anesthetize and kill them.Make frozen sections of these fetal brains.Some sections were stained using conventional HE method,to observe the pathological changes under the light microscope.Detect MCMV early antigen in the brain tissue by immunohistochemistry staining and immunofluorescence assay.Results The survival rates of the infected group were 71.9％.Compared with the control group,intra-amniotic inoculation of MCMV does not affect the rate of fetal survival,fetal absorption,fetal death and the average weight of the heads,but decrease their average weight of the bodies.The pathological changes are found in the brain tissue of the mouse in the infection group.Through enzyme immunohistochemistry assay,there are many MCMV infected cells in brain-ventricular zone,brain subependymal zone,cerebral cortex and hippocampus area in the infection group.Similar findings were observed by immunofluorescence method.Conclusion By intra-amniotic injection of MCMV suspension,murine model of MCMV congenital infection can be successfully established.This model could be used to study the mechanisms of encephalodysplasia caused by congenital CMV infection in vivo.

OBJECTIVEThe effect of allitridin on the transcription levels of immediate-early (ie), early(e) and late (1) genes of human cytomegalovirus (HCMV) was investigated in order to explore the mechanism of allitridin against HCMV.

METHODEstablished the models of HCMV AD169 strain infected cells and AD169 strain infected cells treated with allitridin (9.6 mg x L(-1)), and they were compared with the appropriate dose(2.3 mg x L(-1)) of ganciclovir (GCV). All groups of cells were infected at 2.5 multiplicity of infection (MOI), using SYBR Green real-time PCR method to detect the dynamic change of ul122, ul123, ul54 and ul83 mRNA expression at 0.5, 2, 4, 6, 12, 24 h post-infection.

RESULTThe mRNA levels of ul122 and ul123 in AD169 infected cells treated with allitridin at all time points were markedly lower than those of AD169 infected controls (P<0.05), but there were no significant difference of ul122 gene in AD169 infected cells treated with GCV and AD169 infected cells at 0.5-6 h post-infection. The inhibitory rates of allitridin to AD169 ul122 and ul123 mRNA reached 75.2% and 70.4% at 24 h post-infection, respectively. The expression of ul54 mRNA in two drug-treatment groups at all time points were lower than those of AD169 infected cells group (P<0.05). The inhibitory rates of alltridin and GCV to AD169 ul54 mRNA were 45.4% and 27.2% at 24 h post-infection,respectively. The expression of HCMV ul83 mRNA in all groups rapidly increased after 6 h of infection,which is most obvious in AD169 infected cells group. The inhibitory rates of alltridin and GCV to AD169 ul83 mRNA were 45.9% and 26.2% at 24 h post-infection, respectively.

CONCLUSIONAllitridin could effectively suppress the transcription of ie genes (ul122 and ul123) of HCMV AD169 strain, led the expression of mRNA significantly lowerd. It was able to supress the transcription of egene (ul54) and l gene (ul83) too, indicating that HCMV ie genes may be the key target of allitridin against HCMV.

Allyl Compounds ; pharmacology ; Antiviral Agents ; pharmacology ; Cell Line ; Cytomegalovirus ; drug effects ; genetics ; Genes, Immediate-Early ; genetics ; Genes, Viral ; genetics ; Humans ; Sulfides ; pharmacology ; Transcription, Genetic ; drug effects