Objective To Analyze the immunity to secondary infection in the elderly with severe acute pancreatitis.Methods Totally 105 old patients were included in the present study.The ratio of CD4+ to CD8+,and serum levels of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6),interleukin-10 (IL-10),and interleukin-4 (IL-4) were determined by flow cytometer analysis or ELISA within 3 days after diagnosis of secondary infection.Results Among 105 old patients,58 cases experienced secondary infection.At 3th day after severe acute pancreatitis,the levels of TNF-α,IL-6,IL-4,IL-10,CD4+,CD8+ and CD4+/ CD8+ were (81.3±5.5)ng/L,(141.2±13.7)ng/L,(61.1±7.4) ng/L,(153.8 ±15.2) ng/L,(43.5±5.5)％,(20.7±2.9)％ and (2.4±0.3) in infection group,as compared with those of (50.8±4.7)ng/1,(81.4±11.7)ng/L,(30.8±7.8)ng/L,(100.3± 13.8)ng/L,(31.6±4.6)ng/L,(29.7±3.5)and (1.1±0.4)in control group,respectively,with statistically significant differences betwveen the two groups (t=7.30,6.51,4.87,4.52,2.88,3.41,4.26,all P＜0.05).At 28th day after SAP,the levels of TNF-α,IL-6,IL-4,IL-10,CD4+,CD8 ± and CD4+ /CD8+ were (29.3±5.8)ng/L,(51.7±7.9)ng/L,(33.8±5.1)ng/L,(82.6±9.5)ng/L,(22.1±3.3)％,(47.1±4.3)％ and (0.6±0.3) in infection group,as compared with those of (44.4±5.5)ng/L,(82.2±7.1)ng/L,(65.3±5.5)ng/L,(109.1±9.5)ng/L,(40.5±2.7)ng/L,(33.4±4.5)ng/L and (1.8±0.4) in control group,respectively,showing statistically significant differences between the two groups(t=3.26,4.93,7.32,3.43,7.41,3.81,4.33,all P＜0.05).Conclusions An early excessive immune response and subsequent immune injury is closely related to secondary severe acute pancreatitis.

Objective To investigate changes and the significance of Th17/Treg immune imbalance in secondary systemic infection in patients with severeacute pancreatitis.Methods We selected 21 patients with severe acute pancreatitis and secondary systemic infection (infection group),25 patients with severe alone (non-infection group),20 healthy cases undergoing annual health checkup (control group) in this study.The expression levels of Th17/Treg cells and related cytokines were compared between groups.Results There were significant differences in mortality rate and duration of ICU stay between infection group and non-infection group [23.8％ vs.4.0％,(11.3±3.4) d vs.(7.5±2.8) d,x2=3.949,t=2.890,P=0.047 and0.045].The percentages of Th17 cell andTreg cell,Th17/Treg ratio,mRNA expressions of IL-6,IL-17,IL-23,TGF-β and orphan receptor γt were higher in infection and non infection groups than in control group [(26.4 ± 1.2) ％,(12.8 ± 0.9)％ vs.(3.1±0.8) ％;(6.7±1.6)％,(4.2±1.3)％ vs.(1.3±0.4)％;(4.3±1.0)％,(3.2±1.1)％ vs.(2.4±0.9)％;(7.1±0.8)ng/L,(5.3±0.7)ng/L vs.(0.2±0.1)ng/L;(22.9±2.4)ng/L,(15.6±2.8)ng/L vs.(10.3± 1.5)ng/L;(15.7±2.1)ng/L,(10.2± 1.5)ng/L vs.(8.3± 1.4)ng/L;(23.6±2.2)ng/L,(16.3±1.7)ng/L vs.(11.6±1.1)ng/L;(0.052±0.014),(0.035± 0.010) vs.(0.004±0.001);F=15.761,55.745,9.437,102.788,21.038,16.239,36.957,23.924,respectively,P=0.555,0.000,0.014,0.000,0.002,0.004,0.000,0.000].The mRNA expressions of IL-10 and Foxp3-T were lower in infection and non-infection groups than in control group [(6.4±1.1)ng/L,(10.5 ± 2.1) ng/L vs.(15.4±2.0)ng/L;(0.005±0.001),(0.020±0.007) vs.(0.032±0.009),F=18.995 and 20.608,P=0.003 and 0.002].Conclusions The secondary infection can aggravate the Th17 / Treg immune imbalance in patients with severe acute pancreatitis,and extend the ICU hospitalization days.

Objective To compare ultrasound-and endoscopy-guided methods in nasogastrojejunal tube placement in critical ill patients at bedside.Methods A tot al of 95 critical ill patients planed to perform enteral nutrition (EN) were enrolled.They were divided randomly into ultrasound-guided group (48 patients)and endoscopy-guided group (47 patients).The success rate,the incidence of complications and the time they took were compared between two groups.Results Ultrasound guided group was compared to endoscopy guided group with a lower success rate (81.3％ vs 100％,P =0.003),mainly because of the initial 24-stage having a very low success rate (66.7％ vs 100％,P =0.000),but the success rate of the following 24 cases was significantly improved (95.8％ vs 100％,P =0.338).Ultrasound guided group didn 't cause more complications (2.1％ vs 0,P =1.000),and consumed less time [(13.3 ± 2.8)min vs (15.0 ± 1.4) min,P =0.000].Compared to the following 24 cases,the initial 24 cases of ultrasound guided group in nasogastrojejunal tube placement had the lower success rate (66.7％ vs 95.8％,P =0.023),the longer time-consuming [(15.4 ±2.1)rin vs (11.2 ± 1.4) min,P =0.000],and but didnt cause more complications (4.2％ vs 0,P =1.000).The success rate of different diseases [severe acute pancreatitis(SAP) vs cerebrovascular disease] hadn't statistically significant difference (94.9％ vs 87.5 ％,P =0.300),but the SAP group consumed less time [(12.6 ± 1.9)min vs (15.5 ± 2.0)min,P =0.000].Conclusions The placement of nasogastrojejunal tube under ultrasound guidance represented a safe,quick and effective method to provide enteral nutrition.

OBJECTIVETo study the pattern of CGG repeat instability within germline cells derived from two male fetuses affected with Fragile X syndrome (FXS).

METHODSThe length and methylation status of CGG repeats within the testes of a fetus carrying a full FXS mutation and another fetus carrying mosaicism FXS mutation were analyzed with Southern blotting and AmplideX FMR1 PCR. Immunohistochemistry was also applied for the measurement of FMR1 protein (FMRP) expression within the testes.

RESULTSFor the fetus carrying the full mutation, Southern blotting analysis of the PCR product has detected an expected band representing the full mutation in its brain and a premutation band of > 160 CGG repeats in its testis. Whereas the pattern of premutation/full mutation in mosaic testis was similar to that in peripheral blood and no sign of contracted fragment was found other than a band of about 160 CGG repeats. Immunohistochemistry assay with a FMRP-specific antibody demonstrated a number of FMRP-positive germ cells, which suggested a contraction from full mutation to premutation alleles.

CONCLUSIONThis study has clarified the instability pattern of CGG repeat and expression of FMRP protein within the testes of fetuses affected with FXS, confirming that the CGG repeat can contract progressively within the germline. The FMRP expression in the testis is consistent with spermatogonium proliferation, and thus the contraction from full mutation to unmethylated premutations may occur for the requirement of FMRP expression during spermatogenesis. The better understanding of FMRP function during germ cell proliferation may elucidate the mechanism underlying the contraction of full FXS mutation in male germline.

Abortion, Eugenic ; Blotting, Southern ; Brain ; embryology ; metabolism ; DNA Methylation ; Fatal Outcome ; Fetus ; cytology ; metabolism ; Fragile X Mental Retardation Protein ; genetics ; metabolism ; Fragile X Syndrome ; diagnosis ; genetics ; Humans ; Immunohistochemistry ; Male ; Mosaicism ; Mutation ; Polymerase Chain Reaction ; Spermatozoa ; metabolism ; Testis ; cytology ; embryology ; metabolism ; Trinucleotide Repeat Expansion ; genetics

Objective:To assess the value of combined chromosomal karyotyping and chromosomal microarray analysis (CMA) and/or copy number variation sequencing (CNV-seq) for the prenatal diagnosis for women with advanced maternal ages, and to explore the challenges of prenatal genetic counseling brought by the types of fetal CNVs and uncertainty of related phenotypes.Methods:A retrospective analysis was carried out on 1 841 women with advanced maternal age who underwent interventional prenatal diagnosis at the Prenatal Diagnosis Center of Xiamen University Affiliated Women and Children′s Hospital from January 2017 to December 2020. Routine chromosomal karyotyping analysis and CMA/CNV-seq detection were carried out.Results:CMA/CNV-seq had detected pathogenic variants in 2 cases which had failed karyotyping analysis. Two hundred and twenty one fetal chromosomal abnormalities were detected by karyotyping analysis, among which 187 were detected by CMA/CNV-seq. CMA/CNV-seq analysis of 23 cases with balanced chromosome structural aberrations and 10 cases with low proportion mosaicisms (including a marker chromosome) had yielded a negative result. In addition, 26 cases (26/1 841, 1.4%) with pathogenic CNVs were discovered among those with a normal karyotype, of which 13 (50.0%) were recurrent CNVs associated with neurocognitive impairment, with 22q11.21 microdeletions and microduplications being the most common types (26.92%).Conclusion:The combination of karyotyping analysis and CMA/CNV-seq not only increased the rate of prenatal diagnosis, but also complemented with each other, which has facilitated genetic counseling and formulation of prenatal diagnosis strategy for the affected families.

Objective To compare the CGG-repeat-length and its methylation status in fetal tissues and to explicate the heterogeneity of CGG repeats.Method Multiple tissues from a full mutation (August 2013) and a mosaic aborted fetus of 23-week gestation(May 2012) were collected and genomic DNA from these tissues was extracted.The CGG-repeat-length and methylation status in fetal tissues were determined by a combined strategy of Southern blotting and GC-Rich PCR.FMR1 expression was measured by real time PCR and Western blotting.Result CGG-repeat-length in different tissues of each fetus was similar.A major methylated band in the full mutation range (540 CGG repeats) was detected in the brain,skin,testis and kidney tissues of Case 1.An unmethylated premutation band with 160 CGG repeats,and another two bands with 470 and 1 100 CGG repeats in the full mutation range were shown in the brain,skin,testis,lung,stomach,gut,liver,kidney,heart and blood of Case 2.However,the methylation status of CGG repeats in the mosaic fetus was heterogeneous among different tissues.The lowest premutation ratio was in the brain of the mosaic fetus compared with other tissues,and correspondingly FMR1 expression in its brain was minimum.Conclusion This study clarify the tissue heterogeneity of CGG repeats and provides information for the genetic counseling and clinical diagnosis in fragile X syndrome.Based on the fact that the mosaic fetus' mother is a carrier of full mutation,it is speculated that the maternal CGG repeat has contracted before the differentiation of trilaminar germ disc.

Under the background of high-intensity fattening,probiotics have gradually become wide-ly used feed additives.It can inhibit the proliferation of pathogenic bacteria,maintain the microeco-logical balance in the digestive tract,and improve the host immune function.In order to screen lac-tic acid bacteria strains with potential probiotic effects,23 acid-producing strains were isolated from rumen fluid of healthy Duolang sheep by CaCO3-MRS solid medium.Safe strains were re-screened through primary screening and hemolysis tests.Acid tolerance tests and bile salt tests were used to screen the strains that were relatively adapted to the rumen environment.The survival rate of the strain M2 was 93.80％under pH3.0 and 59.72％under bile salt concentration of 0.30％and the survival rate is higher than other strains under high temperature conditions.Subsequently,morphological observation,molecular biological identification,testing of pathogen antagonism,antibiotic tolerance analysis,growth characteristics detection and other methods were used to fur-ther explore the characteristics of the strain.The results showed that the strain to be tested was Gram-positive bacillus.After 16S rRNA gene sequence comparison,the strain was identified as Lactobac illus salivus,which could be used as a candidate strain to develop probiotics.The research results laid a foundation for its application in Duolang sheep breeding.

Objective: To analyze the effectiveness and response status of China Infectious Disease Automated Alert and Response System (CIDARS) in Hunan province from 2012 to 2016 for improving the system. Methods: To collect the early warning signals, the number of suspected events, the result of on-site investigation, the signal response time and the result of public health emergencies, and the χ² test, correlation analysis and non-parametric test were used to analyze the information on CIDARS in Hunan Province during the period from 2012 to 2016. Results: A total of 108 188 signals were generated by the CIDARS in Hunan Province; The warning involved 30 kinds of infectious diseases and 138 counties (districts), and each county (district) received 3.00 weekly warning messages on average; 100% early warning signal was responded, 2 h response rate was 92.43%; The median response time (P25-P75) was 0.28 (0.11-0.77) h in the single case warning, and the five-year timely response rate showed an upward trend year by year (trend χ²=58.89, P<0.05); the median response time (P25-P75) was 0.56 (0.28~1.06) h in the time series warning, and the five-year timely response rate showed no trend (trend χ²=2.43, P>0.05); the time series warning response was more timely than the single case warning, and the difference between the two timely response rates was statistically significant(χ²=5156.60, P<0.05). The studies showed that the sensitivity, specificity, and detection timeliness of the system were 84.80%, 69.53%, and 2.80 d, respectively; There was no significant difference in the infectious diseases detecting timeliness of CIDARS(H=8.75, P>0.05); the system had the best effect in rubella, and the positive likelihood ratio of rubella is 4.36. Conclusions CIDARS in Hunan province runs well overall with high sensitivity and timely response to the early warning signals, but the specificity of the early warning system needs to be improved. The quality of the epidemic report of infectious diseases needs to be further improved, and the early warning parameters and warning method should be adjusted according to the characteristics of different diseases and the differences among different regions.

Autophagy is an intracellular degradation process that maintains cellular homeostasis. It is essential for protecting organisms from environmental stress. Autophagy can help the host to eliminate invading pathogens, including bacteria, viruses, fungi, and parasites. However, pathogens have evolved multiple strategies to interfere with autophagic signaling pathways or inhibit the fusion of autophagosomes with lysosomes to form autolysosomes. Moreover, host cell matrix degradation by different types of autophagy can be used for the proliferation and reproduction of pathogens. Thus, determining the roles and mechanisms of autophagy during pathogen infections will promote understanding of the mechanisms of pathogen‍‒‍host interactions and provide new strategies for the treatment of infectious diseases.
Autophagy ; Bacteria ; Host-Pathogen Interactions ; Lysosomes ; Signal Transduction

Congenital heart disease (CHD) is the most common birth defect worldwide. Long non-coding RNAs (lncRNAs) have been implicated in many diseases. However, their involvement in CHD is not well understood. This study aimed to investigate the role of dysregulated lncRNAs in CHD. We used Gene Expression Omnibus data mining, bioinformatics analysis, and analysis of clinical tissue samples and observed that the novel lncRNA SAP30-2:1 with unknown function was significantly downregulated in damaged cardiac tissues from patients with CHD. Knockdown of lncRNA SAP30-2:1 inhibited the proliferation of human embryonic kidney and AC16 cells and decreased the expression of heart and neural crest derivatives expressed 2 (HAND2). Moreover, lncRNA SAP30-2:1 was associated with HAND2 by RNA immunoprecipitation. Overall, these results suggest that lncRNA SAP30-2:1 may be involved in heart development through affecting cell proliferation via targeting HAND2 and may thus represent a novel therapeutic target for CHD.
Basic Helix-Loop-Helix Transcription Factors ; Cell Proliferation ; Heart Defects, Congenital/genetics* ; Histone Deacetylases ; Humans ; RNA, Long Noncoding/genetics* ; Transcription Factors