TLR4 mediates I/R injury involving endogenous ligands.Interaction of TLR4 with endogenous ligands provides a critical link between tissue damage and activation of the innate immune response.In the early phase of liver,kidney,heart,or lung I/R injury,endogenous ligands are secreted from several kinds of cells,they are recognized by TLR4.Interaction of TLR4 with endogenous ligands,such as HMGB1,seems to be the most important trigger of inflammation and initiates signaling cascades leading to inflammatory and immune responses.Blocking the interaction of TLR4 with endogenous ligands may be useful in clinical management of inflammation and cellular necrosis caused by ischemic insults.

Objective: Our aims were to classify mycoplasma pneumoniae (MP) clinical isolated strain according to endonuclease map of the PCR-products of the MP P1 protein gene and to investigate MP epidemic state in Shenyang.Methods:The P1 protein gene fragment of MP isolated strain was amplified by using PCR method ,and its products were digested with Hea III, 1.5 % argarose gel electrophoresis.Results:Isolated strain showed the same map as MP FH standard strain.Conclusion: Clinical isolated strain of mycoplasma pneumoniae in Shenyang belongs to groupⅡ.

Objective To investigate the effect of intestinal lymphatic duct ligation and ω-3 polyun saturated fatty acids on intestinal and distant organ in intestinal ischemia-reperfusion injury. Methods Totally 40Sprague-Dawley (SD) male rats (SPF grade)after gastrostomy were equally randomized into sham group (Sham), enteral nutrition (EN) group, enteral nutrition and lymphatic duct ligation (EN + L) group, ω-3 polyunsaturated fatty acids (ω-3PUFA) group, and ω-3PUFA and lymphatic duct ligation (ω-3PUFA + L) group. After 7 days of nutritional intervention, rats were subjected to 60 minutes of intestinal ischemia, ischemia plus mesenteric lymph duct ligation, or sham procedures. After 3 days of continuous nutrition intervention using the original nutrient, lymph nodes, lung, intestine, liver, and blood specimens were harvested. Intestinal permeability and morphology, results of bacterial cultures, and serum cytokines were observed or detected. Result After 3 days of intestinal ischemia-reperfusion (I/R), the body weights of rats in EN group significantly decreased when com pared with the pre-I/R levels (P ＜ 0.05), while the body weights of rats in EN + L group were significantly lower than those in ω-PUFA group and ω-PUFA + L group (P ＜ 0. 05). After one day of intestinal ischemia-reperfusion (I/R), the L/M significantly increased in each group (P ＜0.05 or P ＜0. 01). After 3 days of intestinal ischemiareperfusion (I/R) , the L/M were significantly lower than the level one day after ischemia- reperfusion in EN + L group, ω-PUFA group, and ω-PUFA + L group (P ＜ 0.05). The L/M in EN group and EN + L group were significantly higher than that in ω-PUFA + L group (P ＜ 0. 05). The mucosa thickness and villus height of jejunum in ω-PUFA group and ω-PUFA + L group were significantly higher than those in Sham group, EN group, and EN + L group (P ＜ 0. 01 or P ＜ 0. 05). The mucosa thickness and villus height of ileum in ω-PUFA group and ω-PUFA +L group were also significantly higher than those in EN group (P ＜ 0.05). In ω-PUFA + L group, the serum endotoxin level and tumor necrosis factor-α level were significantly lower than those in EN group (P ＜ 0.05), interleukin (IL) -6 level was significantly lower than that in the ω-PUFA group (P ＜ 0.05), and IL-1 β level was significantly lower than those in other groups (P ＜ 0. 05). In EN group, the lung cell apoptosis index was significantly higher than those in other groups (P ＜ 0.05)and the levels of inducible nitric oxide synthase (iNOS)and myeloperoxidase (MPO) were significantly higher than those in ω-PUFA + L group (P ＜ 0. 05). The level of iNOS was also significantly higher in EN + L group than that in ω-PUFA + L group (P ＜ 0.05). Conclusions Sixty minutes of intestinal ischemia can cause intestinal injury, intestinal barrier dysfunction, and increased permeability of intestine. After 72 h of reperfusion, the intestinal injury can be partially recovered and the permeability can be lower than the post-ischemia level; however, bacterial endotoxin translocation and lung apoptotic cells still exist. Intestinal lymphatic ligation can alleviate the lung damage, promote repair of intestinal mucosa, reduce endotoxin translocation, and attenuate the systemic inflammatory response. EN added with ω-3PUFA is remarkably superior to conventional EN.

Objective To investigate the change of high mobility group box 1 ( HMGBI ) after intestine ischemia reperfusion (I/R) in rats, compare the effect of drainage of intestine lymph fluid on gut barrier, and ex- plore the possible mechanism of iachemia-reporfusion injury. Methods Thirty-two Sprague-Dawley (SD) rats (SPF grade) were randomly divided into4 groups with 8 rats in each group: blank group, sham group, intestine is-chemia-reperfusion (I/R) group, and intestine ischemia-reperfusion with drainage of intestine lymph fluid (I/R +drainage) group. Indicators of gut barrier function damage, translocation of endotoxin, and change of HMGB1 and cytokines were detected after intestine ischemia-reperfusion injury. Results The gut barrier function damage and levels of endotoxin, HMGBI, tumour necrosis factor-alpha ( TNF-α), interleukin-6 ( IL-6 ), interleukin-1 beta (IL-1β), and soluble intercellular adhesion molecule-1 (sICAM-1) were significantly lower in blank group and sham group than in I/R group and I/R + drainage group ( P < 0. 05 ). Compared with the intestine I/R + drainage group, the levels of endotoxin and cytokines were significantly higher in the intestine I/R group. The level of HMGB1 was slightly higher than that in the intestine I/R + drainage group, but such difference was not statistically significant ( P > 0. 05 ). lmmunohistochemical staining also revealed that the expression of HMGB1 was significant- ly higher in I/R group than in I/R + drainage group. Conclusions Intestine iachemia-reperfusion injury can lead to the injury of intestine mucosal barrier and increase HMGB1 level HMGB1 may deteriorate gut barrier function and increase the leveh of systemic cytokines. Drainage of lymph fluid can block the gut-lymph pathway and thus decrease the levels of endotoxin and cytokines in systemic circulation and attenuate intestine ischemia-reperfusion injury.

Objective To explore the situation of respiratory virus co-infection with EV71 and CA16 in patients with hand,foot and mouse disease(HFMD) ,and analyze the influence of co-infection on clinical aspects.Methods From June to October of 2010,there were 348 patients enrolled in the study,with 248 hospitalization cases and 100 mild outpatients.All the patients were diagnosed as HFMD in Beijing You-an Hospital.The viral RNA from the pharynx swab samples were extracted and reversely transcribed by RT-PCR.All the samples were detected with the EV71 and CA16 by real-time fluorescence quantitative PCR.Twelve kinds of respiratory viruses were detected by a commercial multiplex-PCR method.The PCR products were confirmed by electrophoresis.Chi square test was used in the data analysis.Results Of the 348 HFMD patients,36 subjects were detected as positive for respiratory virus co-infection.In the 248 hospitalization cases,111 cases were positive for EV71 or CA16,with eight cases identified with respiratory virus co-infection(7.2%); the other 137 cases were negative for EV71 and CA16,with eleven cases identified with respiratory virus co-infection(7.4%).There was not significant difference between respiratory virus co-infection and the identification of EV71 /CA16(x2 = 0.059,P ＞ 0.05).In the 100 mild outpatients positive for EV71 or CA16,seventeen cases were identified with respiratory virus co-infection(17%).The rate of respiratory virus co-infection in the mild outpatients was much higher than in the severe hospitalization patients(x2 = 4.830,P＜ 0.05).Among the 111 EV71(+) or CA16(+) inpatients,there were 101 cases diagnosed as severe cases(91.0%); similarly,there were 132 cases diagnosed as severe cases(96.4%) among the 137 EV71(-) CA16(-) cases.There was not difference between the identification of EV71/ CA 16 and illness of HFMD(x2 = 3.099,P ＞ 0.05).The leading respiratory virus being identified were HRV A/B,PIV3 and FLU A in the 348 HFMD patients.Conclusions Co-infection with respiratory virus exists in the HFMD patients. However,the respiratory virus infection has no significant influence to the state of HFMD illness.

Objective:To investigate the effect of applying narrative therapy theory into game on improving the quality of bowel preparation in children, and to provide a basis for selecting nursing intervention methods before bowel preparation.Methods:This study was a quasi experimental study. Totally 62 children who took polyethylene glycol electrolyte dispersion for bowel preparation in Gansu Maternal and Child Health Hospital from January to December 2021 were included in this retrospective study. They were divided into control group with 28 cases and experimental group with 34 cases according to random number table method. The control group was given conventional bowel preparation and medication guidance, and the experimental group was given the intervention during bowel preparation by applying narrative therapy theory implanted games. The bowel preparation adequacy rate, complete medication taking rate and parental satisfaction of the two groups were observed and compared.Results:The bowel preparation adequacy rate and complete medication taking rate in the experimental group were 94.12% (32/34) and 52.94% (18/34), respectively, which were higher than 46.43% (13/28) and 10.71% (3/28) in the control group, and the differences were statistically significant ( χ2 = 15.23, 10.41, both P<0.01). According to the average BBPS score, the experimental group had better intestinal cleanliness compared to the control group (7.65 ± 1.07 vs 6.07 ± 1.41, t = -4.87, P<0.01), the difference was statistically significant. 97.06% (33/34) of the parents in the experimental group expressed satisfaction, which was higher than 64.29% (18/28) in the control group, and the difference was statistically significant ( χ2 = 12.74, P<0.05). Conclusions:By applying the narrative therapy theory implanted games, the complete taking of high-dose bowel cleasing agent can be promoted and the quality of bowel preparation can be improved, which is worth suggesting as a nursing intervention method for bowel preparation.

Objective: To establish quantitative real-time PCR (qPCR) method based on Taqman probe for detecting Ekpoma virus (EKV). Methods: According to the conserved region of gene in EKV genome from GenBank, primers and probe for qPCR were designed. Validity and sensitivity were evaluated in this study. Both whole blood and serum of a returnee from Angola were tested by the established EKV-1 and EKV-2 qPCR method . Results: Sensitivity of EKV-1 and EKV-2 qPCR method was respectively 41 copies/μl and 70 copies/μl. Coefficient of variance (CV) was respectively 1.27%, 0.20%, 0.82%; 2.12%, 1.74%, and 1.40%. EKV-2 gene was detected in both whole blood and serum of a returnee from Angola. Conclusions The first EKV-2 gene was confirmed in both whole blood and serum of a returnee from Angola by real-time RT-PCR..

The quality control process throughout the Ebola virus nucleic acid detection in Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab) was described in detail, in order to comprehensively display the scientific, rigorous, accurate and efficient practice in detection of Ebola virus of first batch detection team in SLE-CHN Biosafety Lab. Firstly, the key points of laboratory quality control system was described, including the managements and organizing, quality control documents and information management, instrument, reagents and supplies, assessment, facilities design and space allocation, laboratory maintenance and biosecurity. Secondly, the application of quality control methods in the whole process of the Ebola virus detection, including before the test, during the test and after the test, was analyzed. The excellent and professional laboratory staffs, the implementation of humanized management are the cornerstone of the success; High-level biological safety protection is the premise for effective quality control and completion of Ebola virus detection tasks. And professional logistics is prerequisite for launching the laboratory diagnosis of Ebola virus. The establishment and running of SLE-CHN Biosafety Lab has landmark significance for the friendship between Sierra Leone and China, and the lab becomes the most important base for Ebola virus laboratory testing in Sierra Leone.
China ; Ebolavirus ; classification ; genetics ; isolation & purification ; Hemorrhagic Fever, Ebola ; diagnosis ; virology ; Humans ; Laboratories ; manpower ; standards ; Laboratory Infection ; Quality Control ; RNA, Viral ; genetics ; Sierra Leone

Objective To examine the expression profile of programmed death-ligand 1 (PD-L1) on lung endothelial or epithelial cells,and to determine the specific role of PD-L1 in mouse model of indirect acute lung injury (i-ALI).Methods Eighty male C57BL/6 mice were randomly divided into two parts (both n =40).The effects of different administration routes on the expression of PD-L1 were observed.The mice in each part were randomly divided into sham,i-ALI,i-ALI+small interfering RNA (siRNA) random sequence control,and i-ALI+PD-L1 siRNA which could specifically inhibit PD-L1 expression groups,with l0 mice in each group.i-ALI was reproduced in a mouse model of hemorrhagic shock in combination with a subsequent cecal ligation and puncture (CLP).In sham group,only bilateral femoral arteries were ligated without catheterization or bleeding,and only cecum was separated but perforation was not ligated.Intravenous or intratracheal delivery of PD-L1 siRNA was performed 2 hours following the resuscitation to suppress the expression of PD-L1 on lung endothelial or epithelial cells.The mice in i-ALI+siRNA random sequence control group were given siRNA random sequence without inhibition effect on PD-L1 expression,and those in sham group and i-ALI group were given 100 μL phosphate buffered saline (PBS).The mice were sacrificed at 24 hours after CLP,and samples of blood,lung tissue and bronchoalveolar lavage fluid (BALF) were harvested.Expressions of PD-L1 were determined with flow cytometry.Cytokines and chemokines in plasma,lung tissue and BALF were determined by enzyme linked immunosorbent assay (ELISA).The protein concentration in plasma and BALF and the activity of myeloperoxidase (MPO) in lung tissue were quantitatively measured.The pathological changes in lung tissue were observed under light microscope.Results ① Compared with sham group,PD-L1 expression on lung endothelial or epithelial cells were significantly elevated in i-ALI group [endothelial cells:(27.88 ± 1.53)％ vs.(19.64 ± 1.03)％,epithelial cells:(58.70 ± 8.21)％ vs.(29.23 ± 3.94)％,both P ＜ 0.05].② Mice received intravenous delivery of liposomal-encapsulated siRNA had significantly lower expression of PD-L1 on lung endothelial cells as compared with that of i-ALI group [(21.37 ± 0.76)％ vs.(27.88 ± 1.53)％,P ＜ 0.05].Intratracheal delivery of naked PD-L1 siRNA mainly inhibited the PD-L1 expression on epithelial cell as compared with that of i-ALI group [(31.23±4.71) ％ vs.(58.70±8.21) ％,P ＜ 0.05].The expression of PD-L1 in pulmonary microvascular endothelial cells or pulmonary epithelial cells of i-ALI mice was not affected by siRNA random sequence.③ PD-L1 silencing on pulmonary endothelial cells induced by intravenous delivery of PD-L1 siRNA led to a lower protein ratio of BALF/plasma [(4.48 ± 0.35) × 10-3 vs.(6.11 ± 0.56) × 10-3,P ＜ 0.05] and a decreased MPO activity in lung tissue (U · μg-1 · min-1:2.48 ± 0.47 vs.4.56 ± 0.52,P ＜ 0.05) as compared with that of i-ALI group.Moreover,inflammatory mediator levels such as interleukin-6 (IL-6),monocyte chemoattractant protein-1 (MCP-1),macrophage inflammatory protein-2 (MIP-2) and tumor necrosis factor-α (TNF-α) in lung tissue or plasma were significantly reduced following PD-L1 suppression on endothelial cells as compared with those of i-ALI group [IL-6 (ng/g):177.4±23.2 vs.287.9±57.3,MCP-1 (ng/g):839.6±91.7 vs.1 395.7±211.9,MIP-2 (ng/g):923.7± 107.3 vs.1 700.9±240.2 in lung tissue;IL-6 (ng/L):950.2±192.7 vs.1 828.2±243.6,TNF-α (ng/L):258.7±29.1 vs.443.0 ± 58.1,MCP-1 (ng/L):2 583.8±302.3 vs.4 328.1 ±416.4,MIP-2 (ng/L):1 512.9± 165.6 vs.2 005.9 ± 85.7 in plasma,all P ＜ 0.05],however,there was no significant change in the levels of inflammatory factors in BALF.It was shown in lung tissue histology that PD-L1 silencing on pulmonary endothelial cells induced by intravenous delivery of PD-L1 siRNA led to lessened pulmonary edema and reduced immune cells emigration.Intratracheal delivery of PD-L1 siRNA for PD-L1 suppression on epithelial cells had minimal effects on protein ratio of BALF/plasma,MPO activity,inflammatory mediator expressions in lung tissue,plasma,and BALF as well as lung tissue histology.Conclusion PD-L1 silencing on endothelial cells but not epithelial cells protected mice against hemorrhagic shock-sepsis induced i-ALI.

OBJECTIVE: To analyze the clinical and genetic features of three patient diagnosed with Kleefstra syndrome. METHODS: Whole exome sequencing (WES) was carried out for the probands and their parents. Suspected variants were validated by Sanger sequencing. Copy number variations (CNV) were detected by CNV-seq and validated by real-time PCR. RESULTS: Proband 1 was found to carry a de novo heterogeneous variant (c.823+1G>T) of the EHMT1 gene, which may affect its expression. Based on the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be pathogenic (PVS1+PS2+PM2). Proband 2 was found to carry a de novo missense variant c.439C>G (p.L147V) of the EHMT1 gene, which was predicted to be likely pathogenic (PS2+PM1+PM2+PP3). Proband 3 was found to carry a heterozygous 520 kb deletion at 9q34.3 by CNV-seq. The deletion has encompassed the whole of the EHMT1 gene. Real-time PCR has detected no CNV of this region in her parents. CONCLUSION Variants of the EHMT1 gene probably underlay the disease in these patients. Genetic testing has provided a basis for their clinical diagnosis.
Chromosome Deletion ; Chromosomes, Human, Pair 9 ; Craniofacial Abnormalities ; DNA Copy Number Variations ; Female ; Genetic Testing ; Heart Defects, Congenital ; Humans ; Intellectual Disability/genetics* ; Mutation