ObjectiveTo investigate the expression level of HBV cccDNA in patients in the convalescence stage of hepatitis B virus-related acute-on-chronic liver failure （HBV-ACLF） and its correlation with HBV markers and liver histopathological changes. MethodsA total of 30 patients in the convalescence stage of HBV-ACL who were hospitalized in The Ninth Hospital of Nanchang from January 2015 to October 2023 were enrolled as liver failure group， and 9 patients with chronic hepatitis B （CHB）， matched for sex and age， were enrolled as control group. The content of HBV cccDNA in liver tissue was measured， and its correlation with clinical data and laboratory markers was analyzed. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and a one-way analysis of variance or the Kruskal-Wallis H test was used for comparison between multiple groups； the Fisher’s exact test was used for comparison of categorical data between groups. A Spearman correlation analysis was performed. ResultsThe liver failure group had a significantly lower content of HBV cccDNA in liver tissue than the control group （-0.92±0.70 log₁₀ copies/cell vs -0.13±0.91 log₁₀ copies/cell， t=2.761， P=0.009）. In the liver failure group， there was no significant difference in the content of HBV cccDNA in liver tissue between the HBeAg-positive patients and the HBeAg-negative patients （P>0.05）； there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with different grades （G0-G2， G3， and G4） of liver inflammatory activity （P>0.05）； there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with different stages （S0-S2， S3， and S4） of liver fibrosis （P>0.05）； there was no significant difference in the content of HBV cccDNA in liver tissue between the patients with negative HBV DNA and those with positive HBV DNA （P>0.05）. For the liver failure group， the content of HBV cccDNA in liver tissue was positively correlated with the content of HBV DNA in liver tissue （r=0.426， P=0.043） and was not significantly correlated with the content of HBV DNA in serum （P>0.05）. ConclusionThere is a significant reduction in the content of HBV cccDNA in liver tissue in the convalescence stage of HBV-ACLF. HBV cccDNA exists continuously and stably in liver tissue and can better reflect the persistent infection and replication of HBV than HBV DNA in serum and liver tissue.

Objective To investigate the expression of serum Sestrin2 and Toll-like receptor 7(TLR7)in patients with severe pneumonia and sepsis and their predictive value for heart failure.Methods A total of 86 patients with severe pneumonia and sepsis complicated with heart failure(complicated with heart failure group)and 86 patients with severe pneumonia and sepsis(uncomplicated with heart failure group)who were diagnosed and treated in Shanxi Provincial People's Hospital from February 2022 to May 2023 were studied.Another 86 patients who underwent health examinations were included as the control group.Enzyme linked immunosorbent assay(ELISA)method was applied to measure serum levels of Sestrin2 and TLR7.Color doppler echocardiography was applied to measure cardiac function related indicators:left ventricular ejection fraction(LVEF),left ventricular end diastolic diameter(LVEDD),and left ventricular end systolic diameter(LESD).The expression of Sestrin2 and TLR7 levels and cardiac function in three groups was analyzed.Pearson correlation was applied to analyze the relationship between serum levels of Sestrin2 and TLR7 and cardiac function related indicators in patients with heart failure.Receiver operating characteristic(ROC)curve was applied to analyze the predictive value of serum levels of Sestrin2 and TLR7 in patients with severe pneumonia and sepsis complicated with heart failure.Results The serum levels of Sestrin2(11.59±3.31 ng/ml,16.13±3.62 ng/ml),TLR7(48.93±9.52 ng/ml,61.74±10.11 ng/ml),and cardiac function related indicators[LVEDD(53.28±5.76 mm,62.54±6.11 mm)and LESD(38.16±4.38 mm,48.15±5.02 mm)]were higher in uncomplicated heart failure group and complicated heart failure group than those in control group(7.11±2.34 ng/ml,40.12±10.16 ng/ml,44.86±5.02 mm,29.02±4.07 mm),with significant differences(qSestrin2=13.241,26.659,qTLR7=8.224,20.182,qLVEDD=13.824,29.028,qLESD=18.805,39.359,all P＜0.05),and those values in complicated with heart failure group were higher than those in uncomplicated with heart failure group,with significant differences(q=13.418,11.985,15.203,20.554,all P＜0.05).The LVEF(55.43％±6.62％,41.67％±5.84％)index in uncomplicated heart failure group and complicated heart failure group was lower than that in control group(62.75％±7.16％),with significant differences(q=10.344,29.789,all P＜0.05),and the index in complicated with heart failure group was lower than that in uncomplicated with heart failure group,with significant difference(q=19.455,P＜0.05).Pearson correlation analysis showed that serum levels of Sestrin2 and TLR7 in patients with heart failure were negatively correlated with LVEF(r=-0.419,-0.467,all P＜0.05),and positively correlated with LVEDD and LESD(r=0.456,0.419;0.402,0.437,all P＜0.05).Sestrin2 was positively correlated with TLR7(r=0.641,P＜0.05).ROC curve results showed that the area under the curve(AUC)of serum Sestrin2 combined TLR7 for predicting heart failure in severe pneumonia sepsis patients was 0.940,with sensitivity of 74.9％and specificity of 73.3％.Conclusion The serum levels of Sestrin2 and TLR7 in patients with severe pneumonia and sepsis were elevated and may have good predictive value for patients with heart failure.

Objective To examine the cognitive status and influencing factors of spiritual care among neurosurgery nur-ses,providing a basis for developing a spiritual care management plan for neurosurgery nurses.Methods A convenience sam-pling method was used to select 741 neurosurgical nurses from 35 tertiary hospitals across 14 prefecture-level cities in Guangdong Province from October 2022 to March 2023.A self-developed general information questionnaire and the Chinese version of the Spiritual Care-Giving Scale(C-SCGS)were utilized for the questionnaire survey.Results The average cognitive score for spirit-ual care among neurosurgery nurses was 169.07±27.66.Factors influencing the total score and various dimensions of spiritual care cognition included educational background,receipt of spiritual care education,position,number of night shifts,marital sta-tus,and employment type(P＜0.05).Conclusion Neurosurgical nurses'spiritual care cognition is at a moderate level,indi-cating a need for targeted training programs to enhance their understanding and practice of spiritual care based on their status and characteristics.

Objective:To explore chromosomal copy number variations (CNVs) and pregnancy outcomes in fetuses with mild to moderate isolated ventriculomegaly (IVM), but without other indications for invasive prenatal diagnosis.Methods:A retrospective analysis was conducted on clinical data of 215 singleton pregnancies with mild to moderate IVM (lateral ventricular width≥10-<15 mm) who underwent chromosomal microarray analysis (CMA), not indicated by advanced age, high risk in serum screening or abnormal history of pregnancy, at the Fujian Maternity and Child Health Hospital between June 2016 and March 2023. The 215 fetuses were grouped into mild ( n=167) and moderate ( n=48) IVM;unilateral ( n=142) and bilateral ( n=73) IVM; first diagnosis of IVM before 28 weeks ( n=138) and thereafter ( n=77). Anomalies other than IVM were excluded via three-dimensional color Doppler ultrasound examination between 22 and 26 weeks of gestation. Out of these cases, 129 were confirmed by fetal cranial MRI, 191 underwent chromosomal karyotype analysis, and 202 cases received cytomegalovirus DNA quantification test for amniotic fluid. The detection rates of pathogenic CNVs in various groups were compared using Fisher's exact test. Results:Among the 215 fetuses, 11 cases (5.1%) of chromosomal abnormalities were detected through CMA, including one trisomy 21, five pathogenic CNVs, and five CNVs of uncertain clinical significance. Within the pathogenic CNVs, there were two de novo mutations with 16p11.2 microdeletion and one de novo mutation with 16p11.2 microduplication, while one 16p11.2 microduplication and one Xp22.31 microdeletion were inherited maternally. Of the CNVs of uncertain significance, there were two 16p13.11 microduplications, each inherited from a different parent, one paternally and one maternally; meanwhile, family validation was refused in the other three cases with 3p22.1 microdeletion, 3p26.3 microdeletion, and 9q21.33q22.31 microduplication. The detection rate of pathogenic CNVs in the moderate IVM group was higher than that in the mild IVM group [6.3% (3/48) vs. 1.2% (2/167)], but the difference was not statistically significant ( P=0.083). Similarly, no significant difference was found in the detection rate of pathogenic CNVs when comparing the unilateral IVM group [2.1% (3/142)] with the bilateral IVM group [2.7% (2/73)], nor between the group diagnosed with VM before 28 weeks gestation [2.2% (3/138)] and that diagnosed ≥28 weeks [2.6% (2/77)] (both P>0.05). After the exclusion of fetuses with chromosomal pathogenic abnormalities ( n=11), cytomegalovirus infection( n=1), and additional ultrasound anomalies ( n=7), and several cases with missing data intrauterine outcomes were followed up in 169 IVM fetuses, including 104 (61.5%) improved, 60 (35.5%) unchanged, and five (3.0%) progressed. Follow-ups were successful for 194 women, of which eight pregnancies were terminated (including one trisomy 21, four pathogenic CNVs, one fetal cytomegalovirus infection, and two progressed to severe IVM). Among the 186 newborns, one was diagnosed with X-linked ichthyosis, and one child who progressed to severe IVM before born was followed until 20 months of age without notable phenotypic abnormalities. The rest 184 babies, including those with CNVs of uncertain clinical significance, exhibited no developmental abnormalities during follow-up between the ages of three months and six years. Conclusions:For those fetuses with isolated mild to moderate IVM, but without indications for prenatal diagnosis such as advanced maternal age, high risk in serum screening or abnormal history of pregnancy, remain having the risk for chromosomal aberrations, and 16p11.2 microdeletion/microduplication might be a frequent CNV associated with this condition. Aside from those with pathogenic chromosomal aberrations, fetal cytomegalovirus infection, or progressive enlargement of the lateral ventricles, most fetuses with isolated mild to moderate IVM have a good prognosis.

Objective:To investigate the clinical value of isolated fetal echogenic bowel (FEB) as an indicator for invasive prenatal diagnosis.Methods:This retrospective study enrolled 183 pregnant women who were diagnosed with isolated FEB and underwent invasive prenatal diagnosis in Fujian Maternity and Child Health Hospital from August 2013 to January 2021. Clinical data including the results of conventional karyotyping and chromosomal microarray analysis (CMA), cytomegalovirus (CMV) DNA loads in amniotic fluid and pregnancy outcomes were reviewed analyzed. Chi-square test was used for statistical analysis Results:Karyotyping was performed on all of the 183 fetuses and three (1.64%) aneuploidies (one case of trisomy 21, one trisomy 18 and one 47,XYY syndrome) were detected. One trisomy 21 and four pathogenic (P)/likely pathogenic (LP) copy number variation (CNV) were detected among 108 fetuses who received CMA. The detection rate of P/LP chromosomal abnormalities by CMA was higher than that by karyotyping, but there was no significant difference between them [4.63% (5/108) vs 0.93% (1/108), χ 2=1.54, P>0.05]. In addition, three cases of variants of uncertain significance (VOUS) were detected by CMA. CMV DNA loads of fetal cells in the amniotic fluid samples of the 166 cases were determined, and only one (0.6%) was positive (CMV DNA up to 7.01×10 6 copies/ml), and no abnormalities were found in karyotype analysis and CMA detection. A total of 176 cases were followed up, and among them only one case of intrauterine infection and seven cases (three aneuploidies and four P/LP CNV) of chromosomal abnormalities were terminated after genetic counseling. Three fetuses with VOUS and other 165 fetuses without chromosomal abnormalities had a good prognosis after birth. Conclusions:Isolated FEB may be the abnormal ultrasound finding in fetuses with chromosomal abnormalities or CMV infection. Prenatal genetic testing and the exclusion of intrauterine infection are important for management during pregnancy and prognosis assessment of FEB.

The aim of this study was to develop a semi-quantitative immunochromatographic method for rapid detection of Newcastle disease virus (NDV) antibodies by expressing HN protein in rice endosperm bioreactor. The recombinant plasmid pUC57-HN was digested by MlyⅠ and XhoⅠ to retrieve the HN gene, while the intermediate vector pMP3 containing promoter, signal peptide and terminator was digested by NaeⅠ and XhoⅠ. The HN gene and the linearized pMP3 were purified and ligated to form a recombinant plasmid pMP3-HN1. Subsequently, pMP3-HN1 and plant vector pCAMBIA1300 were digested by EcoRⅠ and Hind Ⅲ, and the HN1 gene was cloned into pCAMBIA1300. The recombinant plasmid pCAMBIA1300-HN1 was introduced into Agrobacterium tumefaciens EHA105 by electrotransformation, and the pCAMBIA1300-HN1 was transferred into rice callus by agrobacterium-mediated method. After dark culture, callus screening, differentiation, rooting and transplanting, transgenic rice seeds were obtained 4 months later. PCR identified that the HN gene has been inserted into the rice genome. SDS-PAGE and Western blotting indicated that the HN protein was successfully expressed in the positive rice endosperm. The purity of the HN protein was more than 90% by SP cation exchange chromatography and gel filtration chromatography. According to the national standards for the diagnostic techniques of Newcastle disease HI test (HI≥4log₂, positive antibody reaction), a colloidal gold labeled purified HN protein was used to prepare a semi-quantitative test strip by double-antibody sandwich method for rapid detection of NDV antibody. The results showed that the test strip did not cross-react with positive sera against other viruses, and the sensitivity of the test strip reached 1:102 400 for standard positive sera of Newcastle disease. Testing of a total of 308 clinical sera showed that the compliance rate of the test strip with HI test was 97.08%, and the Kappa value was 0.942. In conclusion, high purity recombinant HN protein was obtained from rice endosperm, and a simple, rapid, highly sensitive and highly specific semi-quantitative immunochromatographic strip was developed. The test strip could be used for immune evaluation of the Newcastle disease vaccine.
Animals ; Antibodies, Viral ; Chickens ; HN Protein/metabolism* ; Newcastle Disease/prevention & control* ; Newcastle disease virus/metabolism* ; Oryza/genetics*

OBJECTIVE: To assess the impact of confined placental mosaicism (CPM) on non-invasive prenatal testing (NIPT) and pregnancy outcomes. METHODS: Copy number variation sequencing (CNV-seq) and single nucleotide polymorphism array (SNP-array) were carried out on placental specimen sampled from eight pregnancies with confirmed false-positive NIPT results. The impact of CPM on NIPT and pregnancy outcomes were analyzed based on the laboratory tests and clinical characteristics. RESULTS: Five of the eight cases with false-positive NIPT results were proven to be CPM involving trisomy 9, 13, 21, 22, and X, respectively. The mosaic ratios for different placental regions have varied from 4% to 80%. Two fetuses with confirmed CPM showed fetal growth restriction (FGR) and additional ultrasound abnormalities, 1 fetus showed only FGR. The remaining two fetuses showed normal growth. CONCLUSION NIPT is highly sensitive to CPM, whilst CPM is an important cause for false-positive NIPT result. CPM may be associated with FGR. Investigation of the presence of CPM is important for both pre- and post-test genetic counseling and management of the pregnancy.
DNA Copy Number Variations ; Female ; Humans ; Mosaicism ; Pregnancy ; Pregnancy Outcome ; Prenatal Diagnosis ; Trisomy

Objective:To understand the characteristics of the HIV-1 molecular networks in injected drug users (IDUs) in Guangzhou from 2008 to 2017, and provide reference for the prevention and control of AIDS in this population.Methods:The serum samples of newly diagnosed HIV-1 positive IDUs in Guangzhou from 2008 to 2017 were collected, HIV-1 RNA was extracted for pol gene amplification and sequencing. The molecular cluster in the phylogenetic tree was identified by Cluster Picker 1.2.3 for cluster analysis. TN93 model in HyPhy2.2.4 was used to calculate the gene distance between the cluster sequences. Software Cytoscape3.8.2 was used to visualize the molecular network, and χ2 test or exact probability method was used for cluster analysis and centrality analysis. Results:A total of 586 sequences were successfully amplified (73.9%, 586/793), and 80 molecular clusters were produced, with a clustering rate of 46.6% (273/586). In molecular clusters, the proportions of the strains from IDUs in Han ethnic group (48.4%, 260/537), IDUs who were local residents in Guangdong (52.7%, 146/277) and IDUs whose strain sequence subtype was CRF55_01B (93.3%, 14/15) were higher. In the molecular network, the degree range was 1-7, and nodes with degree ≥3 accounted for 12.8% (24/187), which was associated with another 81 nodes in the molecular network (43.3%, 81/187). The centrality analysis showed that the proportions of housework/unemployed with high degree centrality (19.0%, 19/100), high intermediary (22.0%, 22/100), and high proximity centrality (32.0%, 32/100) were higher in IDUs infected with HIV-1.Conclusion:The risk of HIV-1 clustering in IDUs in Guangzhou was high, suggesting that IDUs who were from both Guangdong and Guangxi and were house workers or unemployed should be viewed as the key targets, and precise intervention should be implemented to reduce the HIV-1 infection rate in this population.

Objective To analyze the magnetic resonance imaging (MRI) of the spinal cord and clinical characteristics in patients with autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy.Methods A total of 1 040 samples of cerebrospinal fluid (CSF) and sera collected in the Second Affiliated Hospital of Guangzhou Medical University from March 2013 to June 2018 were tested with tissue-and cell-based assays,and 42 patients were found positive for GFAP-IgG.The clinical data and MRI characteristics of the spinal cord of 19 patients who were positive for GFAP-IgG in CSF with autoimmune GFAP astrocytopathy and lesions in the spinal cord were retrospectively reviewed.Results There were 12 females and seven males among the 19 patients,with onset age of (44±17) years.The main manifestations of these patients included limb weakness (14/19),abnormal vision (5/19),headache (4/19),seizure (4/19),dementia (3/19),etc.On MRI of the spinal cord,five patients showed involvement in the cervical cord alone,eight showed involvement in the thoracic cord alone and six had both cervical and thoracic segment involvement.Fifteen patients had longitudinally extensive myelitic abnormalities (≥3 vertebral segments long).Seven enhancement patterns were encountered.Lesions were displayed in the spinal cord and brain in eight patients.Central gray matter involvement in the spinal cord was found in all the 19 patients.Conclusions Autoimmune GFAP astrocytopathy more frequently presents in females than in males.MRI of the spinal cord has complex presentations and longitudinally extensive myelitic abnormalities usually.Patients often show central gray matter involvement in the spinal cord.Myelitic abnormalities present more often in thoracic segment than in cervical segment.Abnormalities in lumbar segment are less encountered.

OBJECTIVE: To determine the origin of supernumerary small marker chromosomes (sSMCs) carried by two fetuses. METHODS: Single nucleotide polymorphism array (SNP-array) and fluorescence in situ hybridization (FISH) analysis were carried out on cells cultured from the amniotic fluid samples. RESULTS: SNP-array analysis showed both fetuses to be arr[hg19]22q11.1q11.21(16 888 899-18 649 190)×4, with a duplicated 1.7 Mb region (16 888 899-18 649 190) leading to partial tetrasomy of 22q11.1-22q11.21. FISH confirmed that both fetuses were 47,XN,+mar.ish idic(22)(q11.2) (RP11-958H20 ++),which suggested a diagnosis of Cat-eye syndrome (CES). The appearance of abortuses were consistent with the diagnosis of CES. CONCLUSION Two fetuses with CES were diagnosed by genetic testing. The latter has provided a basis for genetic counseling.
Aneuploidy ; Chromosome Disorders ; diagnosis ; Chromosomes, Human, Pair 22 ; Eye Abnormalities ; diagnosis ; Female ; Fetus ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Pregnancy ; Prenatal Diagnosis