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.

Objective To investigate the biological function of M122 in pathogenesis of MCMV in developmental brain disorders and brain damage, screening for mouse brain cDNA library interacting with M122 was performed by a yeast two-hybrid system. Methods The reconstructed bait plasmid pGBKT7-M122 was transformed into yeast cells AH109 and screened on the nutrient deficiency medium SD/-Trp. After express of the bait protein in AH109 yeast strains was detected by Western blot analysis, yeast-two hybrid screening was performed by mating AH109 with Y187 containing mouse brain cDNA library plasmid. The diploid yeast cells were plated on the nutrient deficiency medium SD/-Trp/-Leu/-His/-Ade. The second screening was performed with SD/-Trp/-Leu/-His/-Ade containing X-α-gal. The plasmids in positive colonies were extracted and transformed into E. coli JM109 cells. After plasmid DNA in JM109 cells were extracted form positive colonies and sequenced, the results were analyzed by bioinformatic methods. The interactions between M122 protein and the protein obtained from positive colonies were further confirmed by repeating yeast-two hybrid. Then, autoactivations of the proteins obtained from positive colonies were detected.Results The reconstructed bait plasmid was transformed into yeast cells AH109 successfully. The bait protein expressed in the yeast cells AH109 stably. 24 proteins interacting with MCMV M122 were screened, including syntaxin 8 ( Stx8 ), phosphoglucomutase 2 ( Pgm2 ), potassium voltage-gated channel, shaker-related subfamily, beta member 1 ( Kcnab1 ), collagen, type ⅪⅩ, alpha 1 ( Col19a1 ), archain 1 ( Arcn1 ), cytidylate kinase( Cmpk), DnaJ(Hsp40) homolog, subfamily A, member 1 (Dnaja1), ATPase, Na+/K + transporting, beta 3 polypeptide( Atp1b3 ), SH3-domain GRB2-like ( endophilin ) interacting protein 1 ( Sgip1 ),ankyrin repeat domain 17 (Ankrd17), Smg-7 homolog, nonsense mediated mRNA decay factor(Smg7),sperm associated antigen 9 ( Spag9 ), FK506 binding protein 1a ( Fkbp1a), MYST histone acetyltransferase monocytic leukemia 4 ( Myst4), hyaluronan and proteoglycan link protein 1 ( Hapln1), autophagy-related 3 (Atg3), splicing factor, arginine/serine-rich 5 ( Sfrs5 ), zinc finger, C3HC-type containing 1 ( Zc3hc1 ),thioredoxin-related transmembrane protein 1 ( Txndc1 ), adaptor protein complex AP-1, gamma 1 subunit (Ap1g1), Cullin 1 ( Cul1 ), and so on. Three of them were formerly unknown proteins. M122 protein could interact with the proteins obtained from positive colonies in the yeast cells AH109. Ap1g1 and Cul1 were proved to have autoactivation. Conclusion A class of proteins in brain interacting with M122 has been obtained. It is presumed that these proteins are correlated with neuropathogenesis of the brain disorders caused by CMV, but the candidates still need further confirmation for the interaction.

Objective To determine the utility of diffusion-weighted magnetic resonance imaging ( MR DWI ) in detecting tumor necrosis with histological correlation after neoacljuvant chemotherapy. Methods Conventional MRI and DWI were obtained from 36 patients with histological proven esteosarcoma. Magnetic resonance examinations were performed in all patients before and after 4 cycles of preoperative neoadjuvant chemotherapy. Apparent diffusion coefficients (ADC) were calculated. The degree of tumor necrosis was assessed using the histological Huvos classification after chemotherapy. T-test was performed for testing changes in ADC value between the 2 groups. P value less than 0. 05 were considered as a statistically significant difference. Results The differences in ADC between viable [ (1.06±0. 30) ×10-3mm2/s ] and necrotic [ (2. 39±0. 44 )×10-3mm2/s] tumor were significant (t= 3. 515,P<0. 05). Changes in ADC value was greater in good responses to neoadjuvant chemotherapy than in poor responses, the ADC value in good responses was increased from (1.18±0. 19)×10-3mm2/s to (2. 27±0. 20)×10-3mm2/s, the corresponding value in poor responses was increased from (1.45± 0.11)×10-3mm2/s to (1.83±0. 16)×10-3mm2/s, There was significant difference in changes of ADC values between good responses and poor responses ( t = 4. 981, P < 0. 01 ). Conclusion Diffusion-weighted MRI permits recognition of tumor necrosis induced by chemotherapy in osteosarcoma. DWI is correlated directly with tumor necrosis. They have potential utility in evaluating the preoperative chemotherapy response in patients with primary osteosarcoma.

Aim To observe the variation of T-cell subsets in spleen in different courses of disease and tis-sue histopathological changes in rats with adjuvant ar-thritis (AA).Methods The model of rat AA was in-duced by complete Freund’s adjuvant into the right hind metatarsal footpad of 20 male Lewis rats.Arthritis was evaluated by total score,arthritis index and paw swelling number. The percentage of total (CD3 +CD4 +), memory (CD4 + CD44 +), no-sensitizated (CD4 + CD62L +) and Th1 7 (CD4 + IL-1 7 +) on CD4 +T cells in spleen were detected by flow cytometry on days 1 4 and 22 after immuniozation.The his-topathological evaluation of heart,liver,spleen,lung, kidney,ankle joints and mesenteric lymph nodes were examined by hematoxylin and eosin (HE)stain.Re-sults The onset of secondary arthritis appeared on a-bout day 1 0 after immunization,with a peak onset on day 1 9.Systematic arthritis symptoms included front and hind paw swelling and redness (swelling was more apparent in the front rather than hind paws),nodule formation and redness on the ears,connective tissue swelling and redness of the nose and evident nodules and redness on the tail.Total score,arthritis index and the paw swelling numbers were enhanced in AA rats. Pathohistology of ankles joints showed hyperplasia, pannus formation,and inflammatory cells infiltration in AA rats.And pathohistology of heart,liver,spleen, kidney and mesenteric lymph nodes showed inflamma-tory changes.Compared with normal group,there was no change in the percentage of total,memory and no-sensitizated CD4 +T cell in the early stage inflammation but they were significantly enhanced in the peak of in-flammation.Th1 7 cell subsets were significantly en-hanced in both of the early and peak of inflammation. Conclusion These findings suggest that abnormal T cell responses and tissue histopathological changes are important features of AA rats.

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 sleep characteristics of adult partial epilepsy patients. Methods 45 patients newly diagnosed as adult partial epilepsy and 40 cases of healthy controls were monitored with sleep encephalogram (EEG), the sleep parameters were analyzed.Epworth Sleepiness Scale (ESS) was used to evaluate the daytime sleepiness next day. The sleep parameters and ESS score were compared between 2 groups, and the correlation between sleep parameters and ESS score in the patients were analyzed. Results The ESS score was higher in the patients than in the controls (P<0.05); Compared with the controls, the total sleep time decreased (P<0.05), the sleep efficiency reduced (P<0.05), the percentage of non-rapid eye movement sleep (NREM) stages 1 and 2 and the wakefulness stage increased (P<0.05), the percentage of NREM stages 3 and 4 decreased (P<0.05) in the patients; There was no significant difference in total recording time, sleep latency, REM latency, and arousal times between the patients and the controls (P>0.05). The ESS score was higher in the patients who attacked 3 times a month or more (group A) than less than 3 (group B). Compared with group B, the total sleep time decreased (P<0.05), the sleep efficiency reduced (P<0.05), the percentage of NREM stages 1 and 2 and the wakefulness stage increased (P<0.05), the percentage of NREM stages 3 and 4, and REM decreased (P<0.05) in group A. There was no significant difference in sleep latency, REM latency and arousal times between 2 groups (P>0.05). The ESS score of the patients was negatively correlated with sleep efficiency (r=-0.49, P<0.05) and the percentage of NREM stages 3 and 4 (r=-0.54, P<0.05), and was not correlated with other sleep parameters (P>0.05). Conclusion The sleep architecture of patients with adult partial epilepsy is abnormal, and their daytime sleepiness is more than healthy persons. Epileptic seizure can aggravate sleep architecture abnormality and daytime sleepiness. Abnormal sleep parameters are related with ESS score.

Objective To analyze the distribution features of newly reported cases of occupational welder′s pneumoconiosis (OWP) in Guangdong Province from 2013 to 2022. Methods Data of newly reported cases of OWP from National Occupational Disease Network Direct Reporting System of Guangdong Province from 2013 to 2022 was analyzed using conventional data analyzing Methods. Results A total of 294 cases of OWP were reported in Guangdong Province from 2013 to 2022 with an overall downward trend. The average age at diagnosis for new OWP cases was 44±6 years old, and the median of dust exposure duration was 11 years. Males accounted for 99.3% of the cases, and 94.2% of the cases were phase Ⅰ OWP. Guangzhou City had the highest regional distribution with 74.8% of the cases. The industry distribution was mainly manufacturing, accounting for 86.7% of the cases. The scale of enterprises was most common in small enterprises, followed by large and medium-sized enterprises, accounting for 38.4%, 30.6% and 24.5% of the cases respectively. The economic types of enterprises were mostly private enterprises, followed by state-owned enterprises, accounting for 54.1% and 32.0% of the cases respectively. Conclusion In Guangdong Province, the newly reported OWP cases were clustered in terms of disease stage, regional distribution, industry, enterprise scale and enterprise economic type. There was a tendency of younger age at diagnosis and shorter dust exposure duration.

Objective: To analyze the status of occupational medical examination (OME) institutions in Guangdong Province and the critical control points affecting their future development. Methods: A total of 211 OME institutions registered in Guangdong Province were selected as the research subjects. Their current statue was investigated, and the risk assessment of their future development was carried out based on hazard analysis and critical control point theory. Results: The OME institutions were mainly concentrated in the Pearl River Delta, accounting for 72.0%. The public and private medical and health institutions accounted for 70.6% and 29.4% respectively. The filing rates of different OME categories from high to low in the order were physical factors, chemical factors, dust, other, radiation factors, biological factors (P<0.01), and the filing rates were 89.6%, 88.6%, 84.8%, 63.0%, 7.1% and 1.0%, respectively. The rates of excellence in the field assessment from high to low in the order were hearing atlas analysis personnel, lung function examination operators, chest film reading personnel, chief examination physician, technical director, quality director (P<0.01), and the rates of excellence were 49.4%, 26.2%, 20.6%, 10.8%, 8.2% and 4.6%, respectively. A total of 37 institutions did not submit OME information, of which 23 institutions did not carry out OME work after filing. The required diagnosis rate and confirmed diagnosis rate of suspected occupational diseases in 2020 were 34.3% and 55.3%, respectively. In 2021, the detection rate of suspected occupational diseases was only 1.1‰. Among the institutions with biological monitoring records, 64.1% did not participate in external quality assessment program in laboratory, and the unqualified rate of 38 participating institutions was 55.3%. The risk assessment results show that the future development of OME institutions needs to focus on private institutions, institutions that have not reported OME information, institutions that have missed diagnosis and missed reporting of suspected occupational diseases, institutions that have not participated in external quality assessment program in laboratory and core personnel such as quality managers, technical leaders, and main inspection physicians. Conclusion: OME institutions in Guangdong Province show the characteristics of centralized filing category, centralized filing region and centralized filing institution nature. Combined with the potential risks of OME institutional capacity building, it is necessary to implement classified management, strengthen information construction, supervision and management, implement post responsibilities, and promote the high-quality development of OME institutions.

?? [Abstract]? Objective? To explore the current status of spousal intimacy and role adaptation of neonatals' fathers and to investigate the correlation between their spousal intimacy and role adaptation. Methods? Totally 472 fathers of neonatals born in Yinzhou People's Hospital between January and May 2018 were selected by convenient sampling and investigated with Dyadic Adjustment Scale (DAS) and Father's Role Adaptation Scale (FRAS). The correlation between the two was analyzed. Results? The spousal intimacy score of the 472 fathers of neonatals was (119.34±16.52), and 30.93% of them showed bad spousal intimacy. Their role adaptation score was (75.80±9.03). Pearson correlation analysis showed that spousal intimacy and role adaptation of neonatals' fathers were positively correlated with each other (r=0.307,P＜ 0.05). Conclusions? Fathers' role adaptation process is full of pressure, and good spousal relationship helps to improve the role adaptation levels of neonatals' fathers.