[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.

Objective: To analyze the epidemiological and molecular characteristics of norovirus outbreaks in schools in Xicheng District of Beijing from 2017 to 2022, so as to provide evidence for the prevention and control of norovirus outbreaks in schools. Methods: Data of norovirus outbreaks in schools in Xicheng District, Beijing during 2017 to 2022 were collected and analyzed by descriptive epidemiological methods. Realtime PCR was used to detect the nucleic acid of group GⅠand GⅡnorovirus, the positive norovirus nucleic acid samples were sent to Beijing Center for Disease Control and Prevention for molecular typing. Results: From 2017 to 2022, 185 norovirus outbreaks were reported in schools in Xicheng District, including 166 cluster outbreaks and 19 outbreaks. A total of 2 044 cases were reported, with a total attack rate of 13.92%. There were two peaks in the outbreak time, which were from March to June after the spring semester and from October to December after autumn semester. Primary schools were the most common place of occurrence (101 cases), followed by nursery institutions (68 cases) and secondary schools (16 cases). There were statistically significant differences in the incidence rates among different sites(12.37%, 22.78%, 8.47%, χ2=263.34, P<0.01). There were significant differences in the incidence of vomiting, diarrhea, nausea and stomachache among different students (χ2=263.33, 90.58, 20.42, 30.29, P<0.01). Vomiting was the main symptom in primary school and nursery school children (96.41%, 98.28%), and the diarrhea rate was higher in middle school students (68.22%). The outbreaks were mainly caused by type GⅡ norovirus. The genotype from 2017 to 2021 showed the characteristics of diversity, mainly GⅡ.2[P16], but there was no significant advantage for the GⅡ.2 [P16] during 2019 to 2021. Conclusions The norovirus outbreak in schools in Xicheng district of Beijing from 2017 to 2022 are mainly caused by GⅡ type genome. The main genotype is GⅡ.2[P16]. Norovirus infection mainly occurred in primary schools and kindergartens. For the vulnerable populations, it is necessary to improve the capacity to early identification, student infectious disease management, active infection control and prevention measures, and pathogen surveillance and sporadic case monitoring.

Objective:To compare the prognostic values of different classification by using transpulmonary pressure gradient (TPG), diastolic pressure gradient (DPG) and pulmonary vascular resistance (PVR) in patients with pulmonary hypertension due to left heart disease (PH-LHD), and investigated hemodynamic and clinical factors associated with mortality in patients with PH-LHD.Methods:This was a single-center prospective cohort study. In-hospital patients diagnosed with PH-LHD via right heart catheterization at the Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University, from September 2013 to December 2019 were enrolled. Patients were divided according to TPG (cutoff value 12 mmHg; 1 mmHg=0.133 kPa), DPG (cutoff value 7 mmHg), PVR (cutoff value 3 Wood Units), and the combination of TPG and PVR. Baseline characteristic was recorded. All patients were followed up until the occurrence of endpoint event, defined as all-cause death that occurred during the follow-up period, or until April 18, 2022. Receiver operating characteristic curves were used to compare the predictive value of 3 classification methods for all-cause death in PH-LHD patients. The optimal cutoff values were calculated using Jorden index. Survival analysis was performed using Kaplan-Meier analysis, and log-rank test was used to compare the predictive efficacy of classification methods based on optimal cutoff values or guidance-recommended thresholds for the survival of PH-LHD patients. Variables showing statistical significance in the univariate analysis were incorporated into multivariate Cox regression model to analyze the independent risk factors for all-cause mortality.Results:A total of 243 patients were enrolled, aged (54.9±12.7) years old, including 169 (69.5%) males. During a median follow-up of 57 months, there were 101 (41.6%) deaths occurred. Grouping results were as follows: (1) TPG: TPG≤12 mmHg group 115 patients, TPG>12 mmHg group 128 patients; (2) DPG: DPG<7 mmHg group 193 patients, DPG≥7 mmHg group 50 patients; (3) PVR: PVR≤3 Wood Units group 108 patients, PVR>3 Wood Units group 135 patients; (4) TPG and PVR: TPG≤12 mmHg and PVR≤3 Wood Units group 89 patients, TPG>12 mmHg and PVR>3 Wood Units group 109 patients. PVR ( AUC=0. 698,95% CI:0.631-0.766) had better predictive value for all-cause mortality than TPG ( AUC=0.596, 95% CI: 0.523-0.669) and DPG ( AUC=0.526, 95% CI: 0.452-0.601) (all P<0.05). The optimal cutoff values for TPG, DPG, and PVR were13.9 mmHg, 2.8 mmHg, and 3.8 Wood Units, respectively. Kaplan-Meier analysis based on the optimal cutoff values or guidance-recommended thresholds showed that PVR and TPG were the predictors of survival ( P<0.05), while DPG did not showed significance ( P>0.05). Multivariate Cox regression analysis showed that age, PVR and log 2N-terminal pro-B-type natriuretic peptide were independent risk factors for all-cause mortality in PH-LHD patients (all P<0.05). Conclusion:Classification according to PVR was most valuable in predicting all-cause death in PH-LHD patients, while TPG showed moderate predictive ability and DPG had no predictive value.

Objective:To estimate the influence of metabolic parameters in 18F-FDG PET/CT and clinically relevant indicators on the prognosis of patients with cervical cancer. Methods:A total of 174 patients with cervical cancer (age (53.6±11.1) years) who underwent baseline 18F-FDG PET/CT examination in the Affiliated Hospital of Qingdao University from May 2011 to December 2020 were retrospectively collected. Metabolic parameters (metabolic tumor volume of primary lesion (MTV p), total lesion glycolysis of primary lesion (TLG p), MTV sum of total lesions (MTV total) in the whole body, TLG sum of total lesions (TLG total)) and clinical parameters (International Federation of Gynecology and Obstetrics (FIGO) stage, tumor maximum diameter ( Dmax), et al) were collected. Cox regression and Kaplan-Meier method were performed to evaluate the prognostic and predictive values of those parameters. Results:The follow-up time was 6-120 months, during which 52 patients (29.9%, 52/174) developed progression. The 5-year overall survival (OS), progression-free survival (PFS), local control (LC) and distant metastasis-free survival (DMFS) rates were 83.3%(145/174), 70.1%(122/174), 75.3%(131/174) and 82.8%(144/174), respectively. Cox regression showed that FIGO stage and MTV total were independent factors for predicting PFS, OS and LC (hazard ratio ( HR): 1.005-11.605, all P<0.05). FIGO stage and TLG total were independent factors for predicting DMFS ( HR: 1.002-12.258, all P<0.05). Conclusion:MTV total and FIGO stage are effective predictors of patients with cervical squamous cell carcinoma.

Objective:To study the molecular basis for a proband with A subtype B of the ABO blood group and explore the influence of amino acid variant on the activity of glycosyltransferase (GT).Methods:A proband who had presented at the First Affiliated Hospital of Zhengzhou University on July 2, 2020 was selected as the study subject. Serological identification of the ABO blood groups of the proband and her family members were performed by gel card and test tube methods. The ABO gene of the proband was identified by PCR-sequence specific primers (PCR-SSP) and DNA sequencing. A 3D molecular homologous model was constructed to predict the impact of the variant on the stability of α-(1→3)-D-N-acetylgalactosamine transferase (GTA). Results:The red blood cells of the proband, her mother and two younger brothers showed weak agglutination with anti-A and strong agglutination with anti-B. The sera showed 1~2+ agglutination with Ac and no agglutination with Bc. Based on the serological characteristics, the proband was identified as AwB subtype. Pedigree analysis suggested that the variant was inherited from her mother. The blood group of the proband was identified as A223B type by PCR-SSP. ABO gene sequencing analysis showed that the proband has harbored heterozygous variants of c. 297A>G, c. 467C>T, c. 526C>G, c. 657C>T, c. 703G>A, c. 796C>A, c. 803G>C, c. 930G>A and c. 1055insA. Based on the results of clone sequencing, it was speculated that the genotype was ABO* A223/ ABO* B.01. There were c. 467C>T and c. 1055insA variants compared with ABO* A1.01, and c. 1055insA variant compared with ABO* A1.02. Homologous modeling showed that the C-terminal of A223 GT was significantly prolonged, and the local amino acids and hydrogen bond network have changed. Conclusion:Above results revealed the molecular genetics mechanism of A223B subtype. The c. 1055insA variant carried by the proband may affect the enzymatic activity of GTA and ultimately lead to weakening of A antigen.

Objective:To evaluate the effect of wrist-ankle acupuncture on the pharmacodynamics of remimazolam for loss of consciousness in the patients of different genders and ages undergoing painless gastroscopy.Methods:American Society of Anesthesiologists Physical Status classificationⅠor Ⅱ patients, undergoing elective painless gastroscopy, were divided into 4 groups according to gender and age: agedmale (age≥65 yr) group (group AM), agedfemale (age≥65 yr) group (group AF), young male (18 yr≤age <65 yr) group (group YM), and young female (18 yr≤age <65 yr) group (group YF). After 10 min of wrist-ankle acupuncture on 1st and 2nd areas of the bilateralupper limbs of the patient, modified Dixon′s up-and-down sequential experiment was used for the test.The initial dose of intravenous remimazolam was 0.20 mg/kg. Each time the dose increased/decreased by 0.05 mg/kg. If the patient lost consciousness, the next patient received a lower dose of remimazolam, otherwise a higher dose was given in the next patient. Loss of consciousness was defined as MOAA/S score was 0 or 1, and the process was repeated until 7 turning points appeared.The median effective dose (ED 50)and 95% confidence interval of remimazolam were calculated by probit method. Results:The ED 50 and 95% confidence interval of remimazolam were 0.296(0.233-0.376), 0.319(0.262-0.388), 0.323(0.278-0.375) and 0.344(0.285-0.415)mg/kg in AM, AF, YM and YF groups, respectively. There was no significant difference in ED 50 among the four groups ( P>0.05). Conclusions:Under the action of wrist-ankle acupuncture, the pharmacodynamics of remazolam is comparable when used for the patients of different genders and ages undergoing painless gastroscopy.

Objective:To explore the effects of miRNA-373-3p (miR-373-3p) on the proliferation of nephroblastoma G401 cells through targeted regulation of CD44 expression.Methods:Bioinformatic method was used to predict the possible targeted genes of miR-373-3p based on bioinformatic databases including miRDB, miRanda, PITA and DIANA-microT. G401 cells were taken and transfected with miR-373-3p mimic, mimic negative control, miR-373-3p inhibitor or inhibitor negative control, respectively. Cell proliferation ability was detected by using CCK-8 assay. The number of clones was detected by using clone formation assay. The relative expression level of CD44 mRNA was detected by using real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression level of CD44 protein was detected by using Western blotting. The dual luciferase gene reporter assay was carried out in HEK-293T cells to vertify the target gene of miR-373-3p.Results:Bioinformatic analysis indicated that CD44 was a targeted gene of miR-373-3p. After 24 h transfection, the proliferation activity of G401 cells in miR-373-3p mimic group was decreased compared with that in mimic negative control group (all P < 0.05). After 48 h transfection, the proliferation activity of tumor cells in miR-373-3p inhibitor group was increased compared with that inhibitor negative control group (all P < 0.05). The formed number of clones in miR-373-3p mimic group was reduced compared with that in the mimic negative control group (55.3±2.5 vs. 90.7±2.9), and the difference was statistically significant ( t = 14.57, P < 0.01). The formed number of clones in miR-373-3p inhibitor group was more than that in inhibitor negative control group (115.0±2.7 vs. 92.0±2.4), and the difference was statistically significant ( t = 8.86, P < 0.01). The dual-luciferase gene reporter assay showed that CD44 was a direct targeted gene of miR-373-3p. The relative expression levels of CD44 mRNA in miR-373-3P mimic and mimic negative control group were 0.62±0.03 and 1.00±0.01, respectively, and the difference was statistically significant ( t = 11.28, P < 0.01). The relative expression levels of CD44 mRNA in miR-373-3p inhibitor and inhibitor negative control group were 1.31±0.02 and 1.00±0.00, respectively, and the difference was statistically significant ( t = 12.65, P < 0.01). The CD44 protein expression was decreased in miR-373-3p mimic group, while increased in miR-373-3p inhibitor group. Conclusion:miR-373-3p can inhibit tumor cell proliferation by targeting CD44 in nephroblastoma.

Objective:To observe the clinical effect of Traditional Chinese Medicine (TCM) collapsibility combined with pressure therapy in the treatment of eczema.Methods:From January 2021 to August 2021, 60 patients with lower extremity venous ulcers who were treated in Wangjing Hospital, China Academy of Chinese Medical Sciences were selected and randomly divided into the control group (30 patients) and observation group (30 patients). The control group was treated by the routine pressure therapy, and the observation group was by the TCM nursing intervention on the basis of the control group. Both groups were treated for 2 weeks. The visual analogy scale (VAS),chronic Venous Insufficiency Questionnaire (CIVIQ), Self-rating Anxiety Acale (SAS), and Self-rating Depression Scale (SDS) were measured and evaluated at baseline and after treatment.Results:The total response rate was 100.0% (30/30) in the observation group and 86.7% (26/30) in the control group, and the difference was statistically significant ( χ2=4.29, P=0.038). The VAS (2.47±0.68 vs. 2.97±0.14, t=3.95), SAS (40.31±11.93 vs. 55.87±4.53, t=4.73), SDS (44.30±5.38 vs. 47.61±5.33, t=2.39) scores were significantly lower than those in the control group ( P<0.05), CIVIQ score (86.63±2.74 vs. 84.67±3.23, t=2.54) was significantly higher than that of the control group ( P<0.05). Conclusion:TCM combined with pressure therapy can promote the healing of ulcers in patients with eczema, relieve pain, and improve the quality of life of patients.

Objective:To retrospectively analyze the results of prenatal diagnosis for 67 pedigrees affected with Duchenne muscular dystrophy (DMD) from the Central Plain Region of China and explore the optimal diagnostic strategy.Methods:Probands from the 67 pedigrees were subjected to multiplex ligation-dependent probe amplification (MLPA), next-generation sequencing (NGS) and Sanger sequencing to attain the diagnosis. The fetuses were subjected to prenatal diagnosis, and the results were verified with short tandem repeat (STR) analysis.Results:Among the 67 probands, large deletions, duplications and small mutations have accounted for 73.13% (49/67), 10.45% (7/67), and 16.42% (11/67), respectively. A hotspot deletion has involved exons 45 to 52. There were 11 small mutations, among which four were unreported previously. De novo mutations have accounted for 41.80% (28/67), of which 27 were novel large fragment deletions. Prenatal diagnosis was provided to 39 women who have carried the DMD gene mutations, 10 fetuses were identified as male patients and 7 fetuses were female carriers. Among the prenatal diagnosis results of 28 non-carrier pregnancies, one fetus was a male patient with the same mutation site as the proband. Conclusion:For the prenatal diagnosis of DMD, in addition to known pathogenic mutations, deletion of large fragments should also be excluded. Mothers of probands who do not carry the DMD mutation should also undergo prenatal diagnosis to rule out gonadal mosaicism.

OBJECTIVE：To study the effects a nd mechanism of X iaozheng decoction on the proliferation and migration of uterine leiomyoma cells in rat. METHODS ：Female SD rats were randomly divided into normal control group ，model group ， chemical medicine positive control group （Mifepristone tablets ，2.25 mg/kg），TCM positive control group （Guizhi fuling capsules ， 200 mg/kg），Xiaozheng decoction low-dose ，medium-dose and high-dose groups （1.4，2.8，5.6 g/kg），with 15 rats in each group. Except for normal control group ，other groups were given intramuscular injection of estrogen and progesterone to induce uterine leiomyomas model. On the second day after modeling ，rats in administration groups were given relevant medicine intragastrically ； normal control group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 months. After last administration ，the uterus was removed and its morphology was observed ；the uterine coefficient was calculated. Uterine leiomyoma cells or uterine smooth muscle cells were isolated and cultured. The proliferation rate and migration rate of cells were detected by MTT method and cell scratch test ；flow cytometry was used to detect the cell apoptosis rate ；mRNA expression of HMGB 1 were detected by RT-qPCR. The expression of phosphorylated protein kinase B （p-Akt），nuclear factor κB inhibitor α （IκBα）and phosphorylated transforming growth factor β-activated kinase 1（p-TAK1）were detected by ELISA ；the protein expression of HMGB 1，phospholipid 3 kinase（PI3K），p-Akt in cytoplasm and nuclear factor κB p65（NF-κB p65）in nucleus were detected by Western blot assay. RESULTS ：Compared with normal control g roup，the myometrium of the model group was significantly thickened ，the number of uterine smooth muscle cells were significantly increased and the sizes were 83777930。E-mail：jianghua_ouyang@126.com different，the arrangement of muscle fibers in so me areas was disordered，the uterine coefficient ，and the relative expression of HMGB 1 mRNA and protein were increased significantly （P＜ 0.01）. Compared with model group ，the thickening of uterine myometrium and other symptoms were improved to different extents in Xiaozheng decoction groups and positive control groups ；the uterine coefficient ，cell proliferation rate ，migration rate ，mRNA and protein expression of HMGB 1，the expression of p-Akt and IκBα，protein expression of PI 3K and p-Akt （except for Xiaozheng decoction low-dose group ）in cytoplasm were all decreased significantly （P＜0.05 or P＜0.01）；the cell apoptosis rate ，the expression of p-TAK 1，protein expression of NF-κB p65 in nucleus were all increased significantly （P＜0.05 or P＜0.01）. The effects of Xiaozheng decoction showed a dose-dependent trend. CONCLUSIONS ：Xiaozheng decoction can inhibit the proliferation and migration of uterine leiomyoma cells by down-regulating the expression of HMGB 1，PI3K and p-Akt ，up-regulating the expression of NF-κB p65，so as to promote cell apoptosis.