Objective:To investigate the predictive value of heparin binding protein(HBP) combined with pediatric sequential organ failure assessment(pSOFA) in children with sepsis.Methods:Children with sepsis admitted to PICU of Hunan Provincial People's Hospital (the First Affiliated Hospital of Hunan Normal University) from January 2021 to June 2022 were selected as study group,while those who underwent elective surgery for inguinal hernia and assessment of precocious puberty and short stature during the same period were selected as control group.All children with sepsis were divided into sepsis group and septic shock group according to their severity as well as survival group and death group according to prognosis.The study group was monitored for HBP on the 1st,3rd,and 7th day of admission,while the control group was monitored for HBP on the 1st day of admission.Patients in the sepsis group received pSOFA scores immediately after admission.The laboratory results and HBP concentrations were compared between groups，and a joint model was established in combination with pSOFA to observe its predictive performance in sepsis prognosis.Results:A total of 50 children with sepsis were included in study group，including 45 children with sepsis and five children with septic shock.There were 27 males and 23 females，aged 1 month～13 years（median age two years）.There were 7 deaths in this study，including two patients with sepsis and five patients with septic shock.The HBP concentration in the study group was significantly higher than that in the control group on the 1st day，and the HBP concentration in the group gradually decreased with the prolongation of hospital stay.The concentration of HBP on the first day of septic shock group was higher than that of sepsis group，and the difference was statistically significant（ P<0.001）.The concentration of HBP on the 1st day in the sepsis death group was significantly higher than that in the sepsis survival group（ P=0.023）.The receivor operator characteristic curve analysis showed that HBP and pSOFA had good predictive value for the death of children with sepsis，and the joint model of HBP and pSOFA（75.1×pSOFA-0.1×HBP）had the best predictive performance for the death of children with sepsis，but there was no significant difference with the pSOFA. Conclusion:The HBP level significantly increases in children with sepsis，and gradually decreases with the length of hospital stay，and HBP has great value in predicting the outcome of death in children with sepsis，and the combination of pSOFA could improve its predictive ability of death,but not better than pSOFA.

Objective:To investigate the effect of miRNA-3653-3p (miR-3653-3p) on the proliferation and invasion ability of endometrial cancer cells and its related mechanisms.Methods:The data of 356 endometrial cancer patients were downloaded from the OncoLnc database (http://www.oncolnc.org, updated version 2020), and the Kaplan-Meier method was used to analyze the relationship between the expression level of miR-3653-3p and the overall survival of endometrial cancer patients. The miRGator database (https://bio.tools/mirgator_v2.0, updated version 2019) was used to predict the target gene binding to miR-3653-3p. Human endometrial cancer cell lines AN3CA, HEC-1A, HEC-1B, Ishikawa and human normal endometrial epithelial cell line ESC were selected, and the relative expression level of miR-3653-3p was detected by using quantitative real-time fluorescent polymerase chain reaction (qRT-PCR). The cell line with the lowest expression of miR-3653-3p was selected as the research object, which was divided into the negative control group and miR-3653-3p group, and transfected with the control empty vector plasmid and miR-3653-3p overexpression plasmid. CCK-8 method was used to detect the proliferation ability of cells, Transwell method was used to detect the invasion ability of cells, and qRT-PCR and Western blot were used to detect the expression of miR-3653-3p target gene. The effect of miR-3653-3p on the related protein expression of Wnt- β-catenin signaling pathway was detected by using Western blot.Results:Data analysis in the OncoLnc database showed that compared with endometrial cancer patients with low miR-3653-3p expression, patients with high miR-3653-3p expression had better overall survival ( P < 0.01). Compared with human normal endometrial epithelial ESC, the expression levels of miR-3653-3p in endometrial cancer cell lines AN3CA, HEC-1A, HEC-1B, and Ishikawa were all decreased (all P < 0.05), and the relative expression level of miR-3653-3p was the lowest in HEC-1A cells, and HEC-1A cells were selected for subsequent experiments. The result of CCK-8 showed that compared with the negative control group, the ability of HEC-1A cells in the miR-3653-3p group decreased on the 2nd, 3rd, 4th, and 5th days (all P < 0.05). The result of the Transwell chamber invasion test showed that the number of HEC-1A cell invasion after culturing for 26 h in the negative control group and the miR-3653-3p group was (80±11) and (21±4), respectively, and the difference was statistically significant ( t = 5.18, P < 0.01); compared with the negative control group, the number of cell invasion in the miR-3653-3p group decreased. The miRGator database was used to predict that the target gene of miR-3653-3p might be placenta-specific protein 8 (PLAC8). The relative expression levels of PLAC8 mRNA in HEC-1A cells in the negative control group and miR-3653-3p group were (6.26±0.83) and (0.97±0.31), respectively, and the difference was statistically significant ( t = 6.00, P < 0.01); the relative expression level of PLAC8 mRNA in the miR-3653-3p group was lower than that in the negative control group. Compared with the negative control group, the PLAC8 protein of HEC-1A cells decreased, and the expression of Wnt-β-catenin signaling pathway related proteins β-catenin, transforming growth factor β (TGF-β), GSK-3β, and Rac1 decreased in the miR-3653-3p group. Conclusions:miR-3653-3p may inhibit the proliferation and invasion of endometrial cancer cells by regulating the PLAC8-Wnt-β-catenin signaling pathway.

The heterogeneity of exhausted T cells (Tex) is a critical determinant of immune checkpoint blockade therapy efficacy. However, few studies have explored exhausted T cell subpopulations in human cancers. In the present study, we examined samples from two cohorts of 175 patients with head and neck squamous cell cancer (HNSCC) by multiplex immunohistochemistry (mIHC) to investigate two subsets of Tex, CD8⁺PD1⁺TCF1⁺ progenitor exhausted T cells (TCF1⁺Tex^prog) and CD8⁺PD1⁺TCF1^- terminally exhausted T cells (TCF1^-Tex^term). Moreover, fresh tumor samples from 34 patients with HNSCC were examined by flow cytometry and immunohistochemistry to further investigate their properties and cytotoxic capabilities and their correlation with regulatory T cells (Tregs) in the tumor immune microenvironment (TIME). mIHC and flow cytometry analysis showed that TCF1^-Tex^term represented a greater proportion of CD8⁺PD1⁺Tex than TCF1⁺Tex^prog in most patients. TCF1⁺Tex^prog produced abundant TNFα, while TCF1^-Tex^term expressed higher levels of CD103, TIM-3, CTLA-4, and TIGIT. TCF1^-Tex^term exhibited a polyfunctional TNFα⁺GZMB⁺IFNγ⁺ phenotype; and were associated with better overall survival and recurrence-free survival. The results also indicated that larger proportions of TCF1^-Tex^term were accompanied by an increase in the proportion of Tregs. Therefore, it was concluded that TCF1^-Tex^term was the major CD8⁺PD1⁺Tex subset in the HNSCC TIME and that these cells favor patient survival. A high proportion of TCF1^-Tex^term was associated with greater Treg abundance.
CD8-Positive T-Lymphocytes ; Head and Neck Neoplasms/therapy* ; Humans ; Immunotherapy/methods* ; Prognosis ; Programmed Cell Death 1 Receptor ; Squamous Cell Carcinoma of Head and Neck/therapy* ; Tumor Microenvironment ; Tumor Necrosis Factor-alpha

Objective:To realize the bacterial distribution and antibiotic resistance in children with severe pneumonia in this region.Methods:A total of 203 children with severe pneumonia diagnosed in Gansu Provincial People′s Hospital from April 2018 to March 2020 were divided into 0-1, 1-3, 3-7 and 7-14 years old groups.Bronchoalveolar lavage fluid was collected for bacterial culture and identification, and antibiotic susceptibility tests were performed.Results:The positive rate of pathogens was 69.5% (141/203), including 72.3% (102 strains) of Gram-negative bacteria and 30.5%(43 strains)of Gram-positive bacteria.The infection rates were highest in 0-1 years old group and the lowest in 7-14 years old group, which were 45.2%(19/42) and 16.9%(10/59), respectively.The infection rates of Haemophilus influenzae, Escherichia coli and Branhamella catarrhalis in the 1-3 years old group were 30.30%(10/33), 33.33% (11/33), and 21.21% (7/33), respectively, which showed significant differences compared with other groups( P<0.05). The infection rate of Streptococcus pneumoniae in the 0-1 years old group was 42.9%(18/42), which was significantly different compared with other groups ( P<0.001). The resistance rate of Haemophilus influenzae to trimethoprim/sulfamethoxazole was 89.5%(34/38), and the Streptococcus pneumoniae to trimethoprim/sulfamethoxazole and tetracycline were both 82.4%(28/34). The highest antibiotic resistance rate of Escherichia coli was 34.6%(9/26), and the Branhamella catarrhalis to clindamycin was 56.3%(9/16). Conclusion:The dominant bacteria for severe pneumonia in children are Haemophilus influenzae, Streptococcus pneumoniae, Escherichia coli and Branhamella catarrhalis.The bacterial infection rate is highest within 1 year old, but gradually decreases with the increase of age.Haemophilus influenzae and Streptococcus pneumoniae have severe resistance to several antibiotics.

Objective To determine the serum IgM and IgG antibody levels post-COVID-19 vaccination， and provide scientific evidence for COVID-19 antibody response after vaccination. Methods A total of 980 healthy persons were included in Kunming Third People’s Hospital from July through August， 2021， which had been vaccinated with COVID-19 vaccines and then tested for anti-SARS-CoV-2 IgM and IgG antibodies. Results After the COVID-19 vaccination， 469 persons （positive rate， 47.86%） were positive for anti-IgG antibody. Of them， 75 were males with （positive rate， 39.06%）， and the average IgG level was 0.618 （0.180， 2.526） AU·mL^-1［M（Q₁，Q₃）］； 394 were females （50.00%）， and the IgG level was 0.999 （0.305， 3.334） AU·mL^-1. In addition， 53 persons （5.41%） were anti-IgM antibody positive. Of them， 14 were males （positive rate， 7.29%）， and the average IgM level was 0.057 （0.026， 0.195） AU·mL^-1； 39 were females （4.95%）， and the IgM level was 0.047 （0.027， 0.110） AU·mL^-1. The positive rate of anti-IgG antibody was highest in those aged ≤30 years， which was 51.02% in male （n=25） and 55.88% in female （n=133）. The anti-IgG response differed significantly by gender （χ²=7.401， D=0.135 1， P<0.05）， whereas the anti-IgM response was not significantly different （χ²=1.656， P>0.05）. Among the age groups， anti-IgG antibody level was higher in those aged ≤30 and 51-70 years， with 158 （55.05%） and 122 （52.36%） persons， respectively； the average antibody level was 1.209 （0.426， 4.386） AU·mL^-1 and 1.074 （0.191， 7.670） AU·mL^-1， respectively. The differences in the positive rates of IgG and IgM antibodies and the levels of IgG antibodies among different age groups were statistically significant （P<0.05）. Using Kruskal-Wallis test and Spearman correlation analysis， it showed a high correlation between the IgG and IgM antibodies （r=0.836 4， H=64.82， 20.09， P<0.05）. Conclusion The Anti-SARS-CoV-2 IgG antibody remains high six months post-COVID-19 vaccination， while anti-IgM antibody is low. The IgM and IgG response are higher in the young and elderly. The response differs by gender and age， demonstrating a correlation.

Objective:To summarize the clinical characteristics of children with Mycoplasma pneumoniae pneumonia(MPP) complicated with pleural effusion, and explore the effect of mixed adenovirus infection on children with MPP complicated with pleural effusion.Methods:The clinical data of children with MPP complicated with pleural effusion diagnosed in Children′s Medical Center at the First Affiliated Hospital of Hunan Normal University (Hunan Provincial People′s Hospital) from January 2013 to December 2019 were collected.MPP cases were divided into single infection group and mixed infection group according to whether mixing adenovirus infection.The clinical characteristics were compared between two groups.Results:A total of 180 children with MPP complicated with pleural effusion were included, the male to female ratio was 1.22∶1 (99/81), the age was 66.13 (44.35, 83.98) months, and the most common cases were children over 5 years old (55.56%). The length of hospitalization was 9.00 (7.00, 12.00) days.Fever (93.33%) and cough (98.33%) were the most common clinical manifestations, and mild increases in C-reactive protein, erythrocyte sedimentation rate and D-dimer were the most common laboratory results.Among included children, right pleural effusion was the most common (54.44%), bilateral pleural effusion accounted for 26.67%, and left pleural effusion accounted for 18.89%.Compared with single infection group, the mixed infection group had a longer hospital stay, a higher proportion of oxygen intake, a higher proportion of gamma globulin use, and a higher value of lactate dehydrogenase and aspartate aminotransferase.The results of multivariate Logistic regression analysis showed that compared with single infection group, although the mixed infection group had a higher proportion of gamma globulin use (36.54% vs.10.93%, P<0.05), the length of hospital stay, clinical manifestations, laboratory examination, chest CT and fiberoptic bronchoscopy showed no statistically significant difference between two groups. Conclusion:MPP complicated with pleural effusion is more common in children over 5 years old, especially in the right side.Mild increases of C-reactive protein, erythrocyte sedimentation rate, and D-dimer are more common.The clinical features of MPP complicated with pleural effusion are similar between mixed adenovirus infection group and single infection group.

Tertiary lymphoid structures (TLS) are ectopic lymphoid structures in cancers that are largely associated with favourable prognosis. However, the prognostic value of TLSs in oral squamous cell carcinoma (OSCC) is largely unknown, and the association between tumour infiltrating lymphocytes (TILs) and TLSs has been rarely explored in OSCC. In this study, associated markers of TLS, including peripheral node address (PNAd) in high endothelial venules, CD20 in B cells and CD3 in T cells, were examined in 168 OSCC patients, and survival analysis was performed between TLS-positive and TLS-negative cohorts. We detected the presence of TILs by staining CD8+ cytotoxic T cells and CD57+ NK cells as well. TLSs appeared as highly organized structures in 45 (26.8%) cases. TLS-positive patients had a better 5-year overall survival (OS) rate (88.9% vs. 56.1%, P < 0.001) and relapse-free survival (RFS) rate (88.9% vs. 63.4%, P = 0.002). Moreover, the presence of TLS was an independent prognostic factor for both the 5-year OS rate (hazard ratio [HR] = 3.784; 95% confidence interval [CI], 1.498-9.562) and RFS rate (HR = 3.296; 95% CI, 1.279-8.490) in multivariate analysis. Furthermore, a higher density of CD8+ T cells and CD57+ NK cells was found in TLS-positive sections than in TLS-negative counterparts (P < 0.001), and their combination provided a higher predictive accuracy (AUC = 0.730; 95% CI, 0.654-0.805). In conclusion, our results suggest that TLS is an independent positive prognostic factor for OSCC patients. These findings provide a theoretical basis for the future diagnostic and therapeutic value of TLSs in OSCC treatment.

Objective To analyze genetic testing and prenatal diagnosis of two pedigrees with X-linked ichthyosis.Methods Karyotyping,bacterial artificial chromosomes-on-BeadsTM (BoBs),fluorescence in situ hybridization (FISH) and single nucleotide polymorphism array (SNP-array) were used to detect amniotic fluid and peripheral blood specimens of two pedigrees,one with and one without known family history of ichthyosis.Clinical data was collected and analyzed as well.Results (1) The pedigree without known family history:Prenatal BoBs showed that the XC1 probe of fetus Ⅳ-12 was from 0.36 to 0.50,suggesting the presence of microdeletion.SNP-array analysis of gravida Ⅲ-13 showed a 1.68 Mb copy number deletion at Xp22.31 and four missing Online Mendelian Inheritance in Man (OMIM) genes (HDHD1,STS,VCX and PNPLA4).Fetal SNP-array revealed a deletion of arr[hg19] Xp22.31 (6 455 151-8 135 644)× 0,indicating a maternally inherited one.FISH analysis verified the deletion in STS gene in fetus Ⅳ-12,whose karyotype was 46,XY.The gravida's female cousin (Ⅲ-21) and nephew (Ⅳ-14) also had STS gene deletion,which size was the same as that from the gravida and the fetus.Fetus (Ⅳ-12) was delivered at term by cesarean section with normal skin,but an extensive white scales appeared on the abdomen one week after birth and the symptom was aggravated when the weather was dry.The infant was followed up to eight months old and no other clinical symptoms were found.(2) The pedigree with known family history:SNP-array revealed that a 1.2 Mb copy number deletion at Xp22.31 and four missing OMIM genes (HDHD1,STS,VCX and PNPLA4) were detected in pregnant women (Ⅲ-21),proband (Ⅳ-16) and fetus (Ⅳ-17).FISH analysis of the fetus verified the deletion in STS gene.The karyotype of the fetus was 46,XY.Fetus Ⅳ-17 was delivered at term by cesarean section with normal skin,but white scales widely appeared on the abdomen ten days after birth.The infant was followed up to four months old and no other clinical symptoms were found.Conclusion Molecular genetic techniques such as BoBs,FISH and SNP array are used in combination in this study to provide genetic testing and prenatal diagnosis to two XLI pedigrees,which is helpful for clinical diagnosis and genetic counseling.

Objective To explore the clinical value of Z score in assessing coronary artery lesions (CAL) of children with Kawasaki disease. Methods The clinical records of 102 children with Kawasaki disease from January 2012 to December 2016 in Gansu Provincial Hospital were retrospectively analyzed. The internal diameter of left main coronary artery (LMCA) and right coronary artery ( RCA) was measured by echocardiography (ECHO),and the incidence of CAL was preliminarily judged. The Z scores of LMCA and RCA were calculated on the basis of the coronary artery diameter,the age of the children and the body surface area,and the incidence of CAL was judged again. Results A total of 22 cases(21. 6%) of CAL were found in 102 cases by ECHO examination,of which 18 cases(17. 6%) of LMCA lesions,and 22 cases(21. 6%) of RCA lesions. A total of 33 cases(32. 4%) of CAL were found by calculating the Z score of coronary artery, of which 29 cases(28. 4%) of LMCA lesions and 33 cases(32. 4%) of RCA lesions. There was significant difference between two methods for determining LMCA lesions (χ2=3. 35,P＜0. 05),and there was no sig-nificant difference between two methods for determining RCA lesions (χ2=3. 01,P＞0. 05). Z score of coro-nary artery was more accurate to detect the CAL in Kawasaki disease,especially LMCA lesions. A large coro-nary artery aneurysm was found in the patients with the largest Z score by selective coronary angiography. Conclusion The Z score can be more conductive to assess the CAL in children with Kawasaki disease,and the higher the Z score,the more serious the CAL is.

Objective To investigate the correlation between gene polymorphism of cytotoxic T lymphocyte associated antigen-4 (CTLA-4) and Graves disease (GD) in Qinghai Tibetan.Methods Using retrospective analysis methods,totally 130 cases of GD were selected randomly from June 2012 to November 2016 in the People's Hospital of Qinghai Province;meanwhile,110 normal control cases were selected randomly from Qinghai Tibetan.Then the genotype and allele of CTLA-4 were detected by the method of polymerase chain reaction restriction fragment length polymorphism (RFLP-PCR).Results The distribution of CTLA-4 genotype frequencies (AA,AG,GG) was different between normal control cases and GD in Qinghai Tibetan [6.2％ (8/130) vs 26.4％ (29/110),50.0％ (65/130)vs 58.2％ (64/110),43.8％ (57/130) vs 15.4％ (17/110),x2 =32.105,P ＜ 0.05].Allele (A,G) frequencies were compared between GD and control,the differences were statistically significant [31.2％(81/260) vs 55.5％ (122/220),68.8％ (179/260) vs 44.5％ (98/220),x2 =28.834,P ＜ 0.05].Conclusion Polymorphisms of CTLA-4 exon 1 (49A/G)genotype and allele are closely correlated with GD in Qinghai Tibetan.