Objective:To analyze the mutation spectrum and hematological phenotype of hemoglobin subunit delta(HBD) gene among prenatal population in Xiangtan City, and to provide scientific basis for screening and diagnosis of rare and complex hemoglobinopathia.Methods:Prenatal population underwent screening and genetic testing for thalassemia at Xiangtan Maternal and Child Health Hospital from October 2022 to December 2023 were selected as the study subjects, combined with capillary electrophoresis results, further HBD gene sequencing was performed to identify specific genotypes.Results:A total of 5 371 subjects were enrolled, 22 cases of HBD gene mutations were detected, with a mutation carrier rate of 0.41% (22/5 371). Among them, 14 cases were diagnosed with δ thalassemia, 7 cases were δ abnormal hemoglobin, and 1 case was δ thalassemia combined with δ abnormal hemoglobin. There were 7 HBD mutant genotypes were included, with -77(T>C) being the most common, followed by hemoglobin (Hb) A 2-Huadu and CD34(+GGT); accounting for 68.2% (15/22), 9.1% (2/22), 9.1% (2/22). CD34(+GGT) was a newly discovered genotype, and CD7(GAG>TAG) was the first reported genotype in Chinese population. The Hb content, mean corpuscular volume, and mean corpuscular hemoglobin content were normal or nearly normal in 22 carriers of the HBD gene mutation; capillary electrophoresis demonstrated reduced Hb A 2 content in all cases. Conclusions:HBD gene mutations have been detected in the prenatal population in Xiangtan City, with -77(T>C) being the most common mutation genotype. None of the mutation carriers showed anemia.

Objective To investigate the reasons for the inconsistent results between the vertical auto profile(VAP)method and bio-chemical homogeneous phase(BHP)method in detecting plasma low-density lipoprotein cholesterol(LDL-C),and provide experimen-tal basis for the accurate and quantitative detection of plasma LDL-C levels.Methods A total of 360 plasma samples from diabetes mellitus patients combined with carotid plaque admitted to the Department of Endocrinology of Ningbo Yinzhou Hospital of Traditional Chinese Medicine during January,2022 and January,2023 were collected.The LDL-C levels of these samples were detected by the VAP method and BHP method,respectively.The VAP method uses software to automatically calculate the area under the LDL-C curve after centrifugation of the sample as the LDL-C level(LDL-CVAP)and the BHP method directly detects the LDL-C level(LDL-CBHP)by the special surfactant method.360 samples were divided into the consistent group(group A)and inconsistent group(group B)ac-cording to the relative deviation between the LDL-CBHP and LDL-CVAP methods.Group B was further divided into the LDL-CBHP on the high side group(Group B1)and LDL-CBHP on the low side group(Group B2).Groups B1 and B2 were divided into B1-1,B1-2,B1-3 and B2-1 groups based on the degree of relative deviation.The percentages of samples and levels of lipoprotein a cholesterol[Lp(a)-C],intermediate-density lipoprotein cholesterol(IDL-C),Lp(a)-C and IDL-C[Lp(a)-C+IDL-C],very low-density lipo-protein cholesterol(VLDL-C),total cholesterol(TC)and total triglyceride(TG)in each group were compared.Results The LDL-CBHP levels of 360 samples were significantly higher than that of LDL-CVAP(P＜0.01).The percentage of samples in group B was significantly higher than that in group A,and that of group B1 was significantly higher than that of group B2(P＜0.05).The levels of Lp(a)-C,IDL-C and Lp(a)-C+IDL-C in groups B1-1,B1-2,and B1-3 were significantly higher than those in group A(P＜0.01).The relative deviation between LDL-CBHP and LDL-CVAP in 360 samples was significantly positively correlated with the levels of Lp(a)-C,IDL-C,and Lp(a)-C+IDL-C(P＜0.01).The maximum correlation coefficient was found in Lp(a)-C+IDL-C.Conclusion The results of plasma LDL-C in diabetes mellitus patients combined with carotid plaque detected by the BHP method are significantly different from those detected by the VAP method,which mainly shows that the results of the BHP method are on the high side.The higher the level of plasma Lp(a)-C+IDL-C,the greater the relative deviation between the BHP method and VAP method.The reason for the high results of LDL-C detected by the BHP method may be related to the fact that LDL-CBHP contains irremovable Lp(a)-C and cholesterol carried by IDL-C.The VAP method can be used as an accurate method for detecting real LDL-C without Lp(a)-C and IDL-C.

Objective:To investigate the effect and mechanism of the gastric cancer cells-derived exosome miR-382-5p in-duced by Helicobacter pylori(H.pylori)on the autophagy and polarization of macrophages,providing new clues for further elucidating the carcinogenic mechanism of H.pylori.Methods:Ultracentrifugation and exosome extraction kit were used to extract the exosomes re-leased by the H.pylori stimulated group and the blank control group AGS cells cells,then transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA)and Western blot were employed to identify exosomes.qRT-PCR was used to detect the expres-sion of miR-382-5p in H.pylori induced AGS-derived exosomes.miR-382-5p mimic was transfected into THP-1 macrophages,then the expressions of autophagy markers(LC3Ⅱ,p62,and Beclin-1)were evaluated by Western blot,the number of autophagosomes was detected by immunofluorescence.The expression levels of PTEN protein,downstream proteins PI3K,AKT,mTOR and its phosphory-lated proteins p-PI3K,p-AKT,p-mTOR were detected by Western blot.Flow cytometry was used to detect the expression levels of macrophage phenotypic molecules CD206 and HLA-DR.ELISA was used to detect the secretion of cytokines TNF-α,IL-6,IL-10 and Arginase1 in macrophage supernatants.Results:The extracted exosomes were consistent with exosome morphology and highly ex-pressed the surface marker proteins CD9,CD63 and TSG101.Compared with the blank control group,the expression level of exosom-al miR-382-5p in H.pylori-infected group was significantly increased.miR-382-5p mimic transfection resulted in decreased expression of LC3 Ⅱ and Beclin-1 in macrophages,increased expression of P62 and decreased number of autophagosomes.Moreover,the protein expression level of PTEN was significantly decreased in the miR-382-5p mimic transfection group,while the expression levels of p-PI3K,p-AKT and p-mTOR were significantly increased.miR-382-5p mimic transfection also resulted in increased expression of mac-rophage M2 type marker protein CD206 and decreased expression of M1 type marker protein HLA-DR,as well as increased expres-sions of IL-10 and Arginine1,whereas decreased expression of IL-6 and TNF-α.Pretreatment with the pathway inhibitor BEZ235 par-tially reverses the effects of miR-382-5p on macrophage autophagy and polarization.Conclusion:H.pylori-induced gastric cancer cells-derived exosomal miR-382-5p suppresses macrophage autophagy and induces M2 polarization through down-regulation of PTEN ex-pression and activation of the PI3K/AKT/mTOR signaling pathway.

Clinical phenotypes and gene characteristics of a patient diagnosed with Mowat-Wilson syndrome (MWS) with Hirschsprung′s disease (HSCR) and vaginal atresia in the Department of Neonatal Surgery, Beijing Children′s Hospital, Capital Medical University in March 2021 were analyzed retrospectively.The eight-month-old girl was admitted to the hospital with symptoms of constipation for nine days and abdominal distension for two days.Lower digestive tract radiography and rectal mucosa biopsy results suggested HSCR.The child also had specific facial features and motor development delay.Whole exome test showed a de novo heterozygous mutation, ZEB2 gene c. 2761C>T (p.R921*). After laparoscopic-assisted Soave procedure, the child had normal bowel movements, and no surgery-related compli-cations occurred during the follow-up period.The child′s motor development improved after rehabilitation treatment.According to literature review, 2 female cases show similar clinical manifestations to this girl, but the genotypes were different.This patient expands the clinical phenotype of ZEB2 gene pathogenicity.

Objective:To investigate the effect of Fuzheng-Peiyuan recipe assisted modified VAD program on M protein, myeloma cells and immune function in patients with multiple myeloma (MM). Methods:A total of 96 patients with MM from January 2017 to May 2019 in our hospital were selected as the research objects, and they were divided into two groups according to the random number table method, with 48 patients in each group. The control group was given a modified VAD regimen (vincristine + adriamycin + dexamethasone), and the observation group was given Fuzheng-Peiyuan recipe as an auxiliary VAD regimen. Both groups were treated for 3 cycles. The clinical efficacy, TCM syndrome score, bone pain score, blood creatinine, hemoglobin, blood calcium, M protein, myeloma cells, immune function [Interleukin-2 (IL-2), Interferon-gamma (INF-γ), Interleukin-4 (IL-4), Interleukin-10 (IL-10)] levels and adverse reactions of the two groups were recorded and compared before and after treatment. Results:The total effective rate of the observation group was 81.3% (39/48), and the control group was 62.5% (30/48). The difference between the two groups was statistically significant ( χ2=4.174, P=0.041). The scores of TCM syndromes ( t=4.674, 13.328) and bone pain scores ( t=4.505, 11.398) in the observation group were significantly lower than those in the control group ( P<0.01) at 1 and 3 cycles after treatment; SCr ( t=4.452, 10.039), blood calcium ( t=4.578, 4.155) in the observation group were significantly lower than those in the control group ( P<0.01); HbAlc levels ( t=5.290, 8.871) in the observation group was significantly higher than that of the control group ( P<0.01); M protein ( t=11.145, 33.812), myeloma cells ( t=6.415, 19.731) in the observation group were significantly lower than those in the control group ( P<0.01); serum IL-2 ( t=4.922, 8.789), INF-γ ( t=5.610, 8.886) in the observation group were significantly higher than those in the control group ( P<0.01); IL-4 ( t=4.709, 6.784), IL-10 ( t=5.287, 12.823) in the observation group were significantly lower than those in the control group ( P<0.01). During the treatment, the incidence of adverse reactions in the observation group was 41.7% (20/48) and that in the control group was 62.5% (30/48). The difference between the two groups was statistically significant ( χ2=4.174, P=0.041). Conclusion:Fuzheng-Peiyuan recipe assisted in improving the VAD regimen in the treatment of MM has a significant clinical effect, which can relieve the clinical symptoms of patients and reduce the degree of bone pain, and promote the reduction of myeloma cells in patients, improve their immune function, and improve the tolerance of chemotherapy.

Objective:To explore the related factors influencing the length of hospital stay(LOS) of pregnant women with heart disease (PWHD) after cesarean section.Methods:A total of 306 patients with PWHD who underwent cesarean section from January 2012 to March 2019 were collected. Among them, 203 patients had not undergone heart surgery (uncorrected group) and 103 patients who had undergone heart surgery (corrected group) during the same period. Demographic, perioperative and postoperative data were recorded. Predictors associated with postoperative LOS were determined using univariate and multivariate linear regression analysis models.Results:(1) The median LOS after cesarean section in the uncorrected group was 6 days (5-8 days). The results of univariate linear regression analysis showed that 38 parameters had significant impact on LOS ( P<0.05). The results of multivariate linear regression analysis showed that 5 parameters were independent risk factors for prolonged LOS in the uncorrected group; among them, the median LOS in uncorrected group with hypertensive disorders of pregnancy was 3 days longer than that in patients with PWHD alone [7 days (5-8 days) vs 4 days (4-5 days), β=0.195, P=0.001]; the median LOS in uncorrected group with high serum creatinine was 3 days longer than normal patients [7 days (5-13 days) vs 4 days (4-5 days), β=0.145, P=0.015]; the LOS of patients who chose general anesthesia was 2 days longer than that of patients who chose spinal anesthesia [6 days (4-8 days) vs 4 days (4-5 days), β=0.154, P=0.007]; the LOS of patients with postoperative pulmonary infection was 4 days longer than that of patients without pulmonary infection [8 days (5-15 days) vs 4 days (4-5 days), β=0.269, P<0.01]; the LOS of patients who admitted to ICU after surgery was 2 days longer than that not admitted patients [6 days (5-8 days) vs 4 days (4-5 days), β=0.268, P<0.01]. (2) The median LOS after cesarean section in corrected group was 4 days (4-5 days). The results of univariate linear regression analysis showed that 8 parameters had significant impact on the LOS (all P<0.05). The results of multivariate linear regression analysis showed that 2 parameters, which were American Society of Anesthesiologists (ASA) grade ( β=0.198, P=0.028) and intraoperative blood loss ( β=0.285, P=0.003), were the independent risk factors for prolonged LOS in corrected group. Conclusion:Preoperative with hypertensive disorders of pregnancy, preoperative creatinine increase, intraoperative general anesthesia, postoperative pulmonary infection, and postoperative admission to ICU are independent predictors of prolonged LOS in uncorrected patients with PWHD; ASA classification and intraoperative bleeding are independent predictor of prolonged postoperative LOS in patients with corrected PWHD.

Objective:The purpose of this study is to analyze the surveillance data of the Integrated HCV surveillance in Shanghai and provide a scientific basis for HCV's elimination strategies.Methods:Descriptive statistical analysis and multivariate logistic regression analysis were performed using the multi-dimension results of the Integrated HCV surveillance in Shanghai from 2014 to 2019. Data related to reported HCV cases, HCV gene subtypes surveillance, HCV behavioral risk factors surveillance and HCV-antibody testing results of the community-based general population and high-risk/key populations.Results:The reported incidence rate of acute hepatitis C in Shanghai decreased from 2014 to 2019 ( Z=-4.07, P<0.01); meanwhile, the reported incidence rate of chronic hepatitis C met an upward trend ( Z=10.26, P<0.01), with an annual average, reported incidence rates of 0.18 per 100 000 and 8.60 per 100 000, respectively. Seven hundred forty-four blood samples were subtyped with 16 subtypes from 4 genotypes (GT1, GT2, GT3, and GT6). Among above, 1b (324 cases, 43.55%), 3a (121 cases, 16.26%), 3b (111 cases, 14.92%) and 6a (47 cases, 6.32%) were the principal subtypes. The composition of genotypes varied with decreased 1b and increased 3b and 6a. The major risk factors for HCV infection were blood transfusion ( OR=4.18, 95% CI: 2.79-6.27), surgery ( OR=1.63, 95% CI: 1.26-2.12), sharing syringe ( OR=4.18, 95% CI: 2.75-6.34), pedicure ( OR=2.01, 95% CI: 1.54-2.62), sharing razors ( OR=4.09, 95% CI:1.24-13.51), and unsafe beauty practices ( OR=3.15, 95% CI: 2.13-4.65). HCV antibody screening of 11 groups of high-risk/key populations showed that drug users had the highest HCV-antibody positive rate of 18.81% (1 008/5 358). The anti-HCV positive rate of the general population was 0.16% (7/4 268), which was significantly lower than that of high-risk/key populations from the same year, 2.50%(501/20 002) ( χ2=94.04, P<0.01). Conclusions:Shanghai is a low-endemic area of HCV. Constantly carrying out integrated surveillance and analysis is of great value for early identification of HCV infected people and its risk factors, timely adjustment of prevention and control strategies, and eliminating the public health threat of HCV.

Objective:To investigate the effect of situational simulation drill in nurses training on prevention and treatment of venous thromboembolism (VTE).Methods:Totally 120 nurses in 1-3 years from a tertiary general hospital were selected to participate in the training for prevention and treatment of venous thromboembolism. They were divided into intervention group and control group according to the number of single and double packet ward. A total of 62 nurses in the single wards were selected as the intervention group, and a total of 58 nurses in the double wards were selected as the control group. The control group and the intervention group were trained on the prevention and treatment of venous thromboembolism by traditional teaching method and scenario simulation drill, and the effect was evaluated by Kirkpatrick Model.Results:After training, intervention group training object theory examination score, operation assessment score, clinical nursing work assessment score, and nurses ability to scale score (83.79±6.85), (87.00±4.57), (7.90±0.62), (79.03±8.24) respectively, were higher than the control group (80.67±7.90), (84.02±4.70), (7.21±0.72), (70.86±11.59), the score comparison difference was statistically significant ( t values were -3.660-5.670, P < 0.05); The satisfaction of the training object to the teaching plan, teaching content and teaching form in the intervention group was 88.71% (55/62), 87.10% (54/62) and 87.10% (54/62), which were all higher than the control group 74.14% (43/58), 72.41% (42/58) and 68.97% (40/58), with statistically significant differences ( χ2 values were 4.250, 4.038, 5.804, P<0.05). Conclusion:The application of situational simulation drill in the nursing training of prevention and treatment of VTE is helpful to improve the theoretical skills and clinical work ability of nurses in the prevention and treatment of VTE, and to increase the satisfaction of nurses with the training.

Objective:To explore the abnormal sensory manifestations of children with autism spectrum disorder (ASD).Methods:A total of 269 ASD children with sensory symptoms (ASD toddler group: under 3 years old( n=110), and ASD preschool children: 3-6 years old( n=159)) were investigated by self-made children's sensory abnormality questionnaire, and 175 typically developing(TD) children were compared.SPSS 26.0 software was used for statistical analysis, including descriptive statistics, t test and χ 2 test. Results:The incidence of sensory abnormalities in ASD group was 95.17%, which was higher than that in TD group (78.86%; χ 2=28.224, P<0.001), and the incidence of multiple sensory domain symptoms in ASD group was 84.77%, which was higher than that in TD group (63.77%; χ 2= 22.608, P<0.001). The incidence of sensory symptoms in ASD toddler group was 98.18%, which was not statistically different from that in TD toddler group (92.59%)(χ 2=3.210, P=0.073). The incidence of multiple sensory domain symptoms in ASD toddler group was higher (85.19%), which was statistically different from that in TD toddler group (66.00%)(χ 2=7.613, P=0.006). In the four items of sensory symptoms (auditory hyperresponsiveness, auditory sensory seeking, tactile hyperresponsiveness and tactile sensory seeking), the proportion of moderate to severe of the first three items in ASD group were 66.67%, 72.08% and 61.54%, which were higher than those in TD group (46.81%, 56.36% and 38.46%)(χ 2= 9.652, P=0.002, χ 2= 7.822, P=0.005, χ 2= 4.631, P=0.031), but the proportion of moderate to severe of four items in ASD toddler group were not statistically different from that in TD toddler group at all ( P>0.05). The incidence of functional impairment caused by four items of sensory symptoms in ASD group (68.55%, 65.99%, 63.25% and 48.87%) was higher than those in TD group (45.74%, 28.18%, 26.92% and 27.12%) (χ 2= 12.814, P<0.001; χ 2= 40.456, P<0.001; χ 2= 11.429, P=0.001; χ 2= 7.930, P=0.005). The incidence of functional impairment caused by auditory sensory seeking and tactile hyperresponsiveness in ASD toddler group (57.65% and 55.56%) was higher than that in TD toddler group (31.58% and 0) and the incidence of the other two symptoms in these two groups were not statistically different.There was no statistical difference in the incidence of sensory symptoms between ASD toddler group and ASD preschool group, neither the incidence of multiple sensory domain symptoms ( P>0.05). Conclusion:Sensory abnormalities are common in autistic children, and most of them involve two or more sensory domains. Attention should be paid to the occurrence, quantity, degree and functional impairment of their symptoms.

Objective To investigate whether elevated parathyroid hormone (PTH) levels could induce endothelial - to - mesenchymal transition (EndMT) and adipocyte transition in endothelial cells (ECs), and to determine the possible underlying mechanism. Methods (1) A rat model of secondary hyperparathyroidism and chronic kidney disease (CKD) was established. The adiposity in bone marrow was detected by oil red O staining. Immunofluorescence staining was performed to detect the expression and localization of cluster of differentiation 31 (CD31) and fibroblast-specific protein 1 (FSP1). (2) The human umbilical vein ECs were cultured in vitro. Western blotting was performed to detect protein expressions of EndMT-related markers CD31, FSP1 and α-smooth muscle actin (α-SMA) in interference groups with different PTH concentrations (0, 10-11, 10-9, 10-7 mol/L PTH for 48 h) and times (0, 12, 24, 48 h, 10-7 mol/L PTH), as well as the expression of β-catenin in interference groups with different PTH concentrations. The localizations of CD31, FSP1 and β - catenin were observed by cell immunofluorescence. Protein expressions of adipocytes markers peroxisome proliferator - activated receptor-γ (PPAR-γ) and CCAAT/enhancer binding protein-α (C/EBP-α) by Western blotting and the degree of adipogenesis by oil red O staining were detected after transformed ECs were cultured in adipogenic culture medium for one week. Small interfering RNA (siRNA) was performed to silenceβ - catenin expression. ECs were divided into control siRNA group, β - catenin siRNA group, PTH +control siRNA group and PTH+β-catenin siRNA group. Protein expressions of CD31, FSP1 and PPAR-γby Western blotting and the degree of adipogenesis by oil red O staining were determined. Results (1) In vivo, compared with the control, CKD rats had increased adipocytes in bone marrow (P＜0.05), and the co-expression of CD31 and FSP1 in bone marrow ECs. (2) In vitro, PTH significantly inhibited the expression of endothelial marker CD31 and increased the expressions of mesenchymal markers FSP1 and α-SMA in concentration-and time-dependent manners. These indexes in 10-7 mol/L PTH group and 0 mol/L PTH group, in 48 h group and 0 h group showed statistical differences (all P＜0.05). In PTH group ECs with 10-7 mol/L PTH for 48 h showed FSP1 accumulation in the cytoplasm and reduced expressions of CD31, and ECs had higher expressions of PPAR-γ and C/EBP-α as well as the degree of adipogenesis than those in control group (all P＜0.05). Furthermore, PTH enhanced the nuclearβ-catenin protein levels in ECs in concentration-dependent. The expressions of β-catenin in 10-7 mol/L PTH group and 0 mol/L PTH group showed statistical differences (P＜0.05). β - catenin expressed in the cytoplasm in control group, while it enter into the nucleus in PTH group. Compared with those in PTH+control siRNA group, the expressions of CD31 and PPAR-γ as well as the degree of adipogenesis decreased in PTH+β-catenin siRNA group (all P＜0.05), while the expression of FSP1 increased (P＜0.05). Conclusions PTH induces ECs - to - adipocytes transition by the canonical Wnt/β - catenin signaling pathway, which might account for bone loss in CKD. Silenced β - catenin expression can inhibit PTH-induced EndMT and adipogenesis.