Objective To investigate the correlation between the expression of peripheral blood CD8 + T lymphocytes and the severity degree of hand,foot and mouth disease( HFMD)in children infected by enterovirus( EV)71 at different ages,and further to predict the role of the expression of CD8+ T lympho-cytes played in the occurrence of neurological complications infected by EV71. Methods A total of 138 pe-ripheral blood samples derived from the confirmed HFMD cases were collected in the department of infectious disease at Kunming Children′s Hospital between March and September in 2014,including 33 mild cases,45 severe cases,and 60 critical cases. Patient ages were 9 months to 5 years old. Flow cytometry was used to detect the expression of peripheral blood CD8 + T lymphocytes in all of above patients. Results Compared to the reference value of CD8 + T lymphocytes in normal healthy children at different age groups,the percentage of peripheral blood CD8 + T lymphocytes elevated or increased slightly in patients of all ages,except the obvi-ously increasing expression in the age group of ~2 years and decreasing expression in critical cases of the age group of ~5 years. The expression of peripheral blood CD8+ T lymphocytes significantly increased in mild and severe patients but slightly increased in critical patients of the age group of 9 to 15 months,gradually de-creased in the age group of ~2 years and slightly increased in mild and severe cases but decreased in critical patients of the age group of ~5 years. There were significant differences between the patients in mild condi-tion and in severe condition or critical condition respectively within the age group of ~2 years( P﹤0. 05 ), and there were no significant differences in other age groups between different severity of disease. Conclusion
There are correlations between the expression of CD8+ T cells and the severity of HFMD in patients at different ages. Especially,the patient′s condition developing into severe degree is associated with the rapid decreasing of CD8 + T cells in HFMD patients of the age group of ~2 years. CD8+T cells play an important role in antiviral immune response in HFMD patients of ~2 years old.

Objective To detect serum IgG4 and IgE levels in children with allergic asthma and rhinallergosis to provide an im-portant laboratory evidence for its diagnosis ,treatment and prevention.Methods The serum IgG4 and IgE levels were detected in 118 children patients with allergic asthma ,167 children patients with rhinallergosis and 150 healthy children(control group) under-going physical examination in the same period.Results The levels of serum IgG4 and IgE in the allergic asthma group and rhinal-lergosis group were significantly higher than those in the control group ,the difference was statistically significant (P<0.05).In the intra-group comparison of the allergic asthma group and rhinallergosis group ,the positive rate of serum IgG4 was higher than that of serum IgE ,the difference was statistically significant (P<0.05).The positive rate of joint detection of serum IgG4 and IgE in these two groups was higher than that of single index detection ,the difference was statistically significant (P<0.05).The positive rate of serum IgG4 in the allergic asthma group was lower than that in the rhinallergosis group ,the difference was statistically sig-nificant(P<0.05).The positive rate of serum IgE in the allergic asthma group was lower than that in the rhinallergosis group ,but the difference was not statistically significant (P>0.05).Conclusion Serum IgG4 and IgE have a certain clinical significance in the occurrence ,treatment and surveillance of allergic asthma and rhinallergosis in children.

Objective To discuss the application and significance of homocysteine (Hcy),renal function and serum lipid levels in renal transplantation,by testing those from patients after renal transplantation.Methods Hcy,creatinine (Cr),urea nitrogen (BUN),uric acid (UA),total cholesterol (TC),triglyceride (TG),low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) were detected in transplantation group(n=63) and control group(n=60).Serum Hcy,Cr and BUN of transplantation group were continuously monitored before and 1,3,7,14 days after operation,and the relationship between Hcy and renal function before and after renal transplantation were compared.Results Compared with control group,Hcy,Cr and BUN in transplantation group all increased and the difference between two groups was statistically significant (P<0.05).Hcy,Cr and BUN in transplantation group all decreased after renal transplantation and the differences between two groups had statistical significance (P<0.05).Compared with that before surgery,Hcy,Cr and BUN in transplantation group gradually reduced 1,3,7 and 14 days after surgery,and the differences were statistically significant (P<0.05).Cr and BUN were positively correlated with Hcy (r=0.627,P<0.05).TC and LDL-C in transplantation group were higher than that in control group and the differences had statistical significance (P<0.05) while the difference of TG and HDL-C didn′t have statistical significance (P>0.05).Conclusion Hcy,Cr and BUN can be used as monitoring indicators of efficacy after renal transplantation,also which can be used to observe the incidence and severity of hyperlipidemia.

In this study, a neutral desorption-extractive electrospray ionization mass spectrometry ( ND-EESI-MS) method was developed for the direct and rapid detection of dichlorvos ( DDVP) in honey samples without any sample pretreatment procedure. Under the positive ionization mode, the main characteristic parent ion of DDVP was m/z 223 (MW:222) and daughter ions were m/z 109 and m/z127. Under the optimized working conditions, with the signal intensity of m/z 127 as quantitative index, the quantitative information of DDVP residues in honey was acquired effectively. The results showed that the linear range of DDVP for spiked honey was 5-1000 ng/mL (R2=0. 998) with the limit of detection (LOD) of 1. 0 ng/mL (n=3) and the recoveries for the DDVP spiked honey samples at the concentration levels of 10 , 30 and 400 ng/mL were 93 . 0%-103. 0%, with the relative standard deviations (RSDs, n=6) of less than 4. 4%. Meanwhile, for detection of spiked honey with gas chromatography-flame photometric detector ( GC-FPD ) , the linear range was 5-1000 ng/mL (R2=0. 999) with the LOD of 1. 6 ng/mL(n=3), and the recoveries of DDVP at the spiked honey concentration levels of 10 , 30 and 400 ng/mL were 94 . 9%-110 . 3%, with the RSDs of less than 7. 6%.

Objective:To investigate the changes of T lymphocyte subsets and cytokines in children with he-patitis B virus(HBV)-associated glomerulonephritis (HBV-GN), and their relationship with HBV-DNA load.Methods:Forty-one children who was the first diagnosed with HBV-GN in Department of Pediatrics, the People′s Hospital of Gansu Province and Institute of Infectious Diseases, the First Hospital of Lanzhou University from September 2012 to September 2016 were collected as the objects(HBV-GN group). At the same time, the 40 patients with HBV infection (chronic HBV infection, normal liver and kidney function, normal 24-hour proteinuria quantitation, no hematuria under the microscope, no recent symptoms of cold and fever, etc.) were enrolled as the control group.The levels of T lymphocyte subset, tumor necrosis factor α(TNF-α), interferon-γ (IFN-γ), interleukin (IL)-2, IL-4, IL-6, IL-8 and IL-10 in the HBV-GN group and the control group were compared, and the relationship between HBV-DNA and cell factors was farther analyzed.Results:Compared with the control group, the proportions of CD3 + T, CD4 + T lymphocyte and CD4 + /CD8 + ratio decreased in the HBV-GN group(0.632±0.052 vs.0.692±0.047, 0.204±0.050 vs.0.466±0.038, 0.006±0.002 vs.0.017±0.003, t=1.025, 3.342, 5.234, all P<0.05), and the proportions of CD8 + T lymphocyte was significantly higher than that in the control group (0.411±0.023 vs.0.220±0.043, t=4.452, P<0.01). Besides, IL-2 and IFN-γ levels in the HBV-GN group were significantly lower than those in the control group[(23.36±2.55) ng/L vs.(36.33±1.24) ng/L, (19.20±2.18) ng/L vs.(61.25±2.08) ng/L, all P<0.05], and the serum levels of TNF-α, IL-4, IL-6, IL-8, and IL-10 were significantly higher than those in the control group[(19.60±1.46) ng/L vs.( 6.68±2.32) ng/L, (13.65±3.34) ng/L vs.(1.35±1.52) ng/L, (5.57±1.02) ng/L vs.(1.43±0.57) ng/L, (26.32±3.45) ng/L vs.(9.68±2.55) ng/L, (19.82±2.78) ng/L vs.(1.02±0.56) ng/L, all P<0.01]. Moreover, in HBV-GN patients, there was negative correlation between HBV-DNA load and IFN-γ, IL-2( r=-0.985, -0.943, all P<0.05), and positive relationship in HBV-DNA load with TNF-α, IL-4, IL-6, IL-8 and IL -10 levels( r=0.942, 0.966, 0.953, 0.944, 0.963, all P<0.05). Conclusions:There is an CD4 + /CD8 + imbalance and an abnormal level of cell factors in HBV-GN progression.In further HBV-GN treatment, HBV-DNA and the cell factors should be detected simultaneously to dynamically eva-luate the illness change and the clinical curative effect.

Thirty-six puerperas who underwent emergency cesarean section at Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from January 24, 2020 to February 9, 2020, who all wore medical surgical masks, were retrospectively included in this study. Anesthesia management was performed under tertiary medical protection measures. A dedicated anesthesia equipment was separately sterilized. Narcotic drugs were used for one patient only, and disposable medical supplies were used for anesthetic supplies. Contact transmission should be avoided when a neonate required resuscitation, and early isolation and nucleic acid testing were provided for the neonates. The rate of suspected cases of novel coronavirus (2019-nCoV) was 11% , and the rate of clinically diagnosed cases was 17% before surgery. The rate of clinically diagnosed cases of 2019-nCoV was 22%, the rate of confirmed cases was 8%, and the total positive rate of diagnosis was 31% after surgery. The rate of neuraxial anesthesia was 86%, the rate of general anesthesia was 14%, the time of spinal puncture was (15±7) min, the time of tracheal intubation under general anesthesia was (2.1±1.3) min, the operation time was (95±36) min, and blood loss was (276±166) ml. The Apgar score of newborns was 8.8 ± 0.5. There was 1 neonate whose mother was diagnosed as having 2019 novel coronavirous disease after operation, an oropharyngeal swab specimen was obtained at 36 h of birth, and the swab was tested positive for 2019-nCoV by nucleic acid testing. As of February 10, 2020, an anesthesiologist involved in the operation was diagnosed to have infection by 2019-nCoV. In conclusion, diagnosis of 2019 novel coronavirous disease during pregnancy is more difficult, it is necessary to perform anesthesia management for cesarean section under tertiary medical protection. Although the difficulty in anesthesia operation is increased under tertiary medical protection, anesthesiologists can carry out standardized anesthesia management and guarantee the safety of maternal and infants and anesthesiologists themselves as long as they are rigorously trained and adhere to protective protocols.

OBJECTIVETo explore the role of ompT gene in uropathogenic E. coli (UPEC) CFT073 strain in urinary tract infection (UTI).

METHODSAn ompT deletion mutant (COTD) was generated by λ Red recombineering in the UPEC CFT073 strain, which was characterized by PCR and sequencing. C57B/L6 mouse models of acute UTI with the mutant and wild-type strains were established to compare the colonization abilities of the two strains in the bladder. The adhesion of CFT073 mutant to human unthelial 5637 cells was also investigated in vitro.

RESULTSPCR and DNA sequencing confirmed the loss of ompT gene in the mutant COTD. The in vitro adhesion rate of the mutant strain COTD to 5637 cells was (6.7±2.2)%, significantly lower than that of (8.3±1.9)% of the wild-type strain (P<0.05). In the murine models of acute UTI, the mutant strain showed a mean colonization number of about (17±8)×10⁴ cfu, which was significantly lower than that of (7∓2)×10⁵ cfu of the wide-type CFT073 strain (P<0.05).

CONCLUSIONOmpT gene can be involved in the colonization of UPEC in the bladder tissue and plays an important role in the pathogenesis of UPEC-induced UTI.

Animals ; Bacterial Proteins ; genetics ; Cell Line ; Escherichia coli Infections ; microbiology ; Escherichia coli Proteins ; genetics ; Gene Knockout Techniques ; Humans ; Mice ; Mice, Inbred C57BL ; Porins ; genetics ; Urinary Tract Infections ; microbiology ; Uropathogenic Escherichia coli ; genetics

Thirty-six puerperas who underwent emergency cesarean section at Tongji Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology from January 24, 2020 to February 9, 2020, who all wore medical surgical masks, were retrospectively included in this study.Anesthesia management was performed under tertiary medical protection measures.A dedicated anesthesia equipment was separately sterilized.Narcotic drugs were used for one patient only, and disposable medical supplies were used for anesthetic supplies.Contact transmission should be avoided when a neonate required resuscitation, and early isolation and nucleic acid testing were provided for the neonates.The rate of suspected cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was 11% , and the rate of clinically diagnosed cases was 17% before surgery.The rate of clinically diagnosed cases of SARS-CoV-2 was 22%, the rate of confirmed cases was 8%, and the total positive rate of diagnosis was 31% after surgery.The rate of neuraxial anesthesia was 86%, the rate of general anesthesia was 14%, the time of spinal puncture was (15±7) min, the time of tracheal intubation under general anesthesia was (2.1±1.3) min, the operation time was (95±36) min, and blood loss was (276±166) ml.The Apgar score of newborns was 8.8±0.5.There was 1 neonate whose mother was diagnosed as having coronavirus disease 2019 after operation, an oropharyngeal swab specimen was obtained at 36 h of birth, and the swab was tested positive for SARS-CoV-2 by nucleic acid testing.As of February 10, 2020, an anesthesiologist involved in the operation was diagnosed to have infection by SARS-CoV-2.In conclusion, diagnosis of coronavirus disease 2019 during pregnancy is more difficult, it is necessary to perform anesthesia management for cesarean section under tertiary medical protection.Although the difficulty in anesthesia operation is increased under tertiary medical protection, anesthesiologists can carry out standardized anesthesia management and guarantee the safety of maternal and infants and anesthesiologists themselves as long as they are rigorously trained and adhere to protective protocols.

Objective:To study the clinical characteristics and gene mutations in a family with combined inherited antithrombin (AT) and factor Ⅶ (FⅦ) deficiency, and explore the relationship between AT gene, F7 gene mutations and diseases. Methods:Pedigree investigation. Blood samplesand clinical dataswere collected fromthe proband and her family members (a total of 16 people in 3 generations) who admitted to the First Affiliated Hospital of Wenzhou Medical University in November 2018. The prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), antithrombin activity (AT: A), antithrombin antigen (AT: Ag), protein C activity (PC: A), protein S activity (PS: A), FⅦ activity (FⅦ: C), FⅦ antigen (FⅦ: Ag) and other indicators were detectedto confirm the diagnosis. DNA direct sequencing analysis of all exons, flanking sequences, 5′ and 3′ untranslated regions of AT genes and F7 genes, and the mutation sites were confirmed by clone sequencingor reverse sequencing. Results:The AT: A and AT: Ag of the proband were 46% and 135 mg/L, respectively (reference range: 250-360 mg/L), some of her family members′ s (father, aunt, two cousins, younger brother and nephew) AT: A and AT: Ag were reduced to 50% of normal range. Her father (Ⅰ 2), aunt (Ⅰ 4), elder brother (Ⅱ 7), younger brother (Ⅱ 8), and nephew (Ⅲ 3)′s FⅦ: C were 45%, 50%, 48%, 47% and 48%, respectively; and their FⅦ:Ag was within the normal range. Genetic analysis revealed that the proband(Ⅱ 6) and some of her family members (father, aunt, two cousins, younger brother and nephew) took rs3138521 polymorphism in the 5′ untranslated region of AT gene. Her father (Ⅰ 2), aunt (Ⅰ 4), elder brother (Ⅱ 7), younger brother (Ⅱ 8), nephew (Ⅲ 3) took c.1091G>A heterozygous missense mutationin exon 8 of F7 gene, resulting in p.Arg304Gln. Conclusion:The rs3138521 in AT gene and c.1091G>A in F7 gene, which may be the molecular mechanism leading to combined inherited AT and FⅦ deficiency in this family.

OBJECTIVE: To analyze the phenotype and genetic variants of a pedigree affected with hereditary protein C (PC) deficiency. METHODS: The protein C activity (PC:A) of the proband and her family members (a four-generation pedigree including 11 individuals) were tested by chromogenic substrate method, and the protein C antigen (PC:Ag) was detected with an enzyme-linked immunosorbent assay(ELISA). The 9 exons and flanking sequences of the protein C (PROC) gene were amplified by PCR and directly sequenced. Suspected mutation was validated by clone sequencing and in other members of the family. MutationTaster and ClustalX-2.1-win was used to analyze the pathogenicity and conservation of the mutation site,respectively. Three-dimentional protein model and amino acids interaction were analyzed with Swiss-PdbViewer software. RESULTS: The PC: A and PC: Ag of the proband were decreased to 46% (reference range: 70%-130%) and 50% (referencerange:70%-140%), respectively. Her grandmother,aunt, cousin and younger brother also showed declined PC:A and PC:Ag by approximately 50%. Genetic analysis revealed that the above individuals have all carried a deletional mutation c.1212-1212delG (p.Met364TrpfsX15) in exon 9 of the PROC gene which can cause replacement of Methionine at position 364 by Tryptophan and alteration of 15 downstream amino acids, and produce a premature stop codon at position 378. The score of MutationTaster was 1.000, indicating that the variant is pathogenic. Conservative analysis showed that the 15 altered amino acids are located in a conserved region across nine homologous species. Protein model analysis showed that the mutation has disrupted a catalytic domain of protein C thereby affected its function. CONCLUSION The heterozygous c.1212-1212delG deletional mutation in exon 9 of the PROC gene, which was unreported previously,probably accounts for the decrease of PC:A and PC:Ag in this pedigree.