[Objective] To explore the accuracy and feasibility of non-invasive prenatal diagnosis of fetal RhE genotype using cell-free fetal DNA (cff-DNA) from maternal peripheral blood. [Methods] A total of 134 pregnant women with single fetuses and RhE-negative blood group were selected from our hospital from November 2023 to August 2024. Free DNA extraction kit was used to extract free DNA from peripheral blood of pregnant women, and the RhE blood group genotype of free DNA was detected by real-time fluorescent quantitative PCR (RT-qPCR). If the qPCR amplification signal of the sample was negative, the methylated RASSF1A gene was amplified, and the positive amplification result was used as a sign of successful extraction of cff-DNA. Serological microcolumn gel method was used to detect the phenotype of RhE blood group in neonatal peripheral blood. [Results] Among the 134 maternal peripheral blood samples, the cff-DNA detection of RhE blood group phenotypes was consistent with the RhE blood group genotyping of neonatal peripheral blood in 133 cases, including 90 cases of Rhee genotype and 43 cases of RhE genotype, with diagnostic concordance rate of 99.3%, sensitivity of 97.7%, specificity of 100%, youden index of 0.977, area under ROC curve of 0.995, the Kappa value of 0.983, positive predictive value of 100%, and negative predictive value of 98.9%. The sample of 1 case failed to be detected. After the amplification of methylated RASSFIA gene, it was confirmed that the reason for the failure was that no cff-DNA was extracted from the sample. The diagnostic concordance rates of the first, second and third trimesters were 93.8% (15/16), 100% (51/51) and 100% (67/67), respectively. Fisher's exact test method was used to calculate the P value, which was P>0.05, indicating that there was no statistical significance in the difference of diagnostic concordance rate among the three pregnancy periods, and there was no difference in the detection concordance rate of this method in different pregnancy periods. [Conclusion] The use of cff-DNA in maternal peripheral blood for the detection of fetal RhE blood group genotype is an accurate and highly feasible non-invasive prenatal diagnostic method, which is helpful for the clinical diagnosis of fetal and neonatal hemolytic disease caused by anti-E antibody.

To conduct timely surveillance of the seasonal characteristics and disease burden of Human Respiratory Syncytial Virus (HRSV) in various geographical regions of China, and further develop more precise and effective prevention and intervention strategies, there is an urgent need for China to establish a nationwide, effective, and stable HRSV surveillance system. Through combining the current status of domestic and international HRSV surveillance systems and the existing surveillance framework in China, this study proposed an HRSV surveillance type applicable to China based on different surveillance purposes, and considering the feasibility of implementation. This article aimed to provide solid scientific and technical support to monitor the dynamic changes of HRSV epidemic timely, carry out a risk assessment and early warning, and further understand the disease burden of HRSV in China. It also helps to improve the diagnosis, prevention, and control of the HRSV diseases research and development, use, and evaluation of HRSV vaccines and drugs in China.

To conduct timely surveillance of the seasonal characteristics and disease burden of Human Respiratory Syncytial Virus (HRSV) in various geographical regions of China, and further develop more precise and effective prevention and intervention strategies, there is an urgent need for China to establish a nationwide, effective, and stable HRSV surveillance system. Through combining the current status of domestic and international HRSV surveillance systems and the existing surveillance framework in China, this study proposed an HRSV surveillance type applicable to China based on different surveillance purposes, and considering the feasibility of implementation. This article aimed to provide solid scientific and technical support to monitor the dynamic changes of HRSV epidemic timely, carry out a risk assessment and early warning, and further understand the disease burden of HRSV in China. It also helps to improve the diagnosis, prevention, and control of the HRSV diseases research and development, use, and evaluation of HRSV vaccines and drugs in China.

OBJECTIVE: To investigate the effect of RUNX2 gene overexpression vector modified exosomes derived from bone marrow mesenchymal stem cells (BMSCs) combined with calcium carbonate scaffold system in bone defect. METHODS: Rabbit BMSCs were used as the research object, and BMSCs were identified by flow cytometry. Construct RUNX2 gene overexpression vector, transfect BMSCs with lentivirus, and collect exosomes by ultracentrifugation. The morphology of exosomes was observed by transmission electron microscope, the expression of exosome marker CD63 was detected by Western blot, and the calcium carbonate scaffold was constructed by three chamber parallel automatic temperature control reaction system. According to whether the RUNX2 gene overexpression vector was transfected or not, the complex of BMSCs and calcium carbonate scaffold was divided into three groups, namely BMSCs group, RUNX2 overexpression group and exosome group. The osteogenic differentiation of BMSCs was detected by oil red O staining and RT-PCR. There were 9 clean adult healthy male New Zealand white rabbits, aged (12.97±1.21) months, with a body weight of (19.3±3.6) kg, with 3 rabbits in each group. The animal model of skull defect was constructed by surgical method, and the repair of bone defect was evaluated by imaging, he staining and Masson staining. RESULTS: The results of flow cytometry showed that the expression of CD29 protein, CD44 protein, CD11b protein and CD45 protein on the surface of BMSCs were 99.5%, 100%, 0.1% and 0.1%, respectively. Transmission electron microscopy showed that the exosomes were bilayer vesicles with a diameter of 50 to 150 nm. Western blot showed that the molecular marker CD63 of exosomes was positive. Oil red O staining showed that the osteogenic differentiation of BMSCs in exosome group was significantly higher than that in RUNX2 overexpression group and BMSCs group. The results of RT-PCR showed that the relative expressions of RUNX2, BMP-2 and ALP mRNA in BMSCs in exosome group were significantly higher than those in RUNX2 overexpression group and BMSCs group (P<0.05). The imaging results showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group. HE staining and Masson staining showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group (P<0.05). MSCs in exosome group was significantly higher than that in RUNX2 overexpression group and BMSCs group. The results of RT-PCR showed that the relative expressions of RUNX2, BMP-2 and ALP mRNA in BMSCs in exosome group were significantly higher than those in RUNX2 overexpression group and BMSCs group(P<0.05). The imaging results showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group. HE staining and Masson staining showed that the repair effect of skull defect in exosome group was better than that in RUNX2 overexpression group(P<0.05). CONCLUSION Compared with RUNX2 gene overexpression vector transfection, extraction of exosomes directly can promote the differentiation of BMSCs into osteoblasts more efficiently, and the combination with calcium carbonate scaffold can better promote the healing of bone defects. So as to provide new ideas and methods for the clinical treatment of bone defects.
Animals ; Calcium Carbonate/metabolism* ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Exosomes/metabolism* ; Humans ; Male ; Osteogenesis/genetics* ; RNA, Messenger/metabolism* ; Rabbits

OBJECTIVE: To explore the effects of siRNA hsa-circ-0000885 modified bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation, cell proliferation and apoptosis in order to provide new ideas and methods for the clinical treatment of osteoporosis (OP). METHODS: From September 2018 to February 2020, 13 patients with osteoporosis admitted to our hospital were selected as the research objects, including 11 females and 2 males, with an age of (65.45±10.77) years old. After obtaining the informed consent of patients, peripheral blood tissues were extracted. Then the expression level of cir-cRNA in peripheral blood mononuclear cells(PBMC) was detected by circ RNA chip. The expression of circ RNA was silenced by siRNA technology. The BMSCs were transfected with lentivirus. According to the siRNA interference plasmid hsa-circ-0000885, the cells were divided into the blank group, the empty vector group and the siRNA interference group. After 72 hours of treatment, the cell cycle was detected by flow cytometry, the apoptosis level was detected by AV-PI kit, and the osteogenic differentiation ability of BMSCs was detected by ALP staining. RESULTS: The expression of hsa-circ-0000885 in PBMC of patients with osteoporosis was significantly higher than that of healthy controls ( CONCLUSION The lentivirus mediated siRNA hsa-circ-0000885 plasmid transfected into BMSCs and osteoclast co culture system can promote cell proliferation, inhibit apoptosis and promote osteogenic differentiation of BMSCs, which can be used as a potential therapeutic target for OP patients.
Aged ; Apoptosis/genetics* ; Cell Differentiation ; Cell Proliferation/genetics* ; Cells, Cultured ; Coculture Techniques ; Female ; Humans ; Lentivirus ; Leukocytes, Mononuclear ; Mesenchymal Stem Cells ; Middle Aged ; Osteoclasts ; Osteogenesis/genetics* ; RNA, Small Interfering/genetics* ; Transfection

Objective:To explore the association between lifestyle-related factors and colorectal adenoma.Methods:Based on the Screening Project of Early Diagnosis and Treatment of Colorectal Cancer in Jiashan county Zhejiang province, from August 2012 to March 2018, information gathered through records on questionnaire and colonoscopic diagnosis were collected from participants with positive results during the primary screening stage. According to the findings of colonoscopy, 11 232 controls without any colorectal diseases and 3 895 cases with colorectal adenoma were included in the study. Multivariate logistic regression models were used to analyze the association between lifestyle-related factors and colorectal adenoma.Results:After adjusting for possible confounding factors, results from multivariate logistic regression analysis showed that smoking, alcohol drinking and obesity were positively related to the risk of colorectal adenoma, with ORs (95 %CIs) as 1.38 (1.24-1.54), 1.37 (1.24-1.51) and 1.38 (1.20-1.59) respectively. However, regular aspirin intake was negatively related with the risk of colorectal adenoma ( OR=0.65, 95 %CI: 0.53-0.80). After stratified by sex and age, data showed that the associations between smoking, alcohol drinking and colorectal adenoma were statistically significant in males, and the association between regular aspirin intake and colorectal adenoma was also statistically significant in older participants (aged 60 years and older). Conclusion:Smoking, alcohol drinking, regular aspirin intake and obesity were associated with colorectal adenoma.

Objective: To explore the correlation between obesity and the risk of colorectal adenoma, so as to provide theoretic evidence for the intervention of the high-risk population for colorectal cancer. Methods: Based on the Screen Project of Early Diagnosis and Treatment of Colorectal Cancer in Jiashan County, from August 2012 to March 2018, the results of colonoscopy and body measurement information of the high-risk population for colorectal cancer were collected. According to the results of colonoscopy, 3 895 patients with colorectal adenoma and 11 232 healthy controls were enrolled. Multivariate logistic regression was used to analyze the correlation between overweight (body mass index (BMI) 24.0 to 27.9 kg/m²), obesity (BMI≥28.0 kg/m²) and the risk of colorectal adenoma. Results: After adjusting for gender and age, compared with that of individuals with normal weight (BMI 18.5 to 23.9 kg/m²), the risk of colorectal adenoma of obese patients increased by 36% (odds ratio (OR)=1.36, 95% confidence interval (CI) 1.18 to 1.56). After stratifing by gender, compared with that of individuals with normal weight, the risk of colorectal adenoma of obese males increased by 30% (OR=1.30, 95%CI 1.07 to 1.59), the risk of colorectal adenoma of overweight females and obese females increased by 15% (OR=1.15, 95%CI 1.01 to 1.31) and 40% (OR=1.40, 95%CI 1.14 to 1.71), respectively. After stratifing by age, compared with that of individuals with normal weight, the risk of colorectal adenoma of obese patients aged between 40 and 59 years increased by 31% (OR=1.31, 95%CI 1.07 to 1.61), and the risk of colorectal adenoma of overweight and obese patients aged between 60 and 74 years increased by 13% (OR=1.13, 95%CI 1.01 to 1.27) and 39% (OR=1.39, 95%CI 1.15 to 1.70), respectively. The results of subgroup analysis according to pathological types indicated that the risk of non-advanced adenoma and advanced adenoma of obese patients increased by 35% (OR=1.35, 95%CI 1.16 to 1.57) and 39% (OR=1.39, 95%CI 1.06 to 1.83), respectively. Conclusions Obesity is correlated with colorectal adenoma, which is more significant in women, individuals aged between 60 and 74 years and advanced adenoma. The intervention of high-risk population for colorectal cancer should include body mass control.

Objective: To investigate the clinical efficacy of Xiaoer anorexia granule combined with Saccharomyces brassicus in the treatment of infantile anorexia and its effect on serum cytokines. Method: Totally 240 cases of infantile anorexia treated in Jinhua People's Hospital from October 2015 to October 2017 were randomly divided into three groups. Group A (combination group) orally took Xiaoer anorexia granule, 1 pack/time for 1-3 years old, 2 packs/time for 4-6 years old, 3 times a day, and combined with saccharomyces brasiliensis, 0.5 g/time, 2 times a day, 15-30 min before meal. Group B took saccharomyces brasiliensis by the same method as the combination group. Group C took Xiaoer anorexia granules by the same method as above,with 80 cases in each group. 8 weeks is a course of treatment. Serum inflammatory cytokines leptin, SP and Ghrelin were also detected. The clinical efficacy, body weight, height improvement and serum cytokine levels were observed in 3 groups. Result: The total improvement rate of group A was 91.25%, which was significantly higher than 71.25%of group B and 73.75%of group C, with statistically significant differences (P<0.01). The total effective rate of group A was 95.00%, which was significantly higher than 71.25%of group B and 70.005%of group C, with statistically significant differences (P<0.01). After 8 weeks of treatment, the serum leptin level in group A was significantly lower than that in group B and group C (P<0.01). The levels of serum SP and Ghrelin in group A were higher than those in group B and C (P<0.01). However, there was no significant difference in cytokine between group B and group C. The 3 groups of patients who received effective treatment were followed up for 6 months (excluding missing cases). There was no significant difference in body weight and height among three groups before treatment. After treatment, the body weight and height of three groups were higher than those before treatment (P<0.05, P<0.01). However, group A was superior to group B and group C (P<0.01). There was no significant difference between group B and group C. Conclusion: Treatment of infantile anorexia with Xiaoer anorexia granules combined with Saccharomyces cerevisiae can improve the total clinical effective rate, adjust serum cytokine levels and increase height and body mass.

Objective: To study influencing factors for aspirin resistance in women with ischemic stroke.Methods: A total of 150 women with ischemic stroke, who visited to our hospital from Jun 2013 to Jun 2015, were selected.According to presence of aspirin resistance, they were divided into aspirin resistance group (n=61) and aspirin sensitive group (n=89).All patients received aspirin enteric-coated tablets on morning, then arachidonic acid pathway platelet aggregation inhibition rate and glycosylated hemoglobin A1c (HbA1c) level were measured in two groups on 10 h after administration.According to HbA1c level, patients were divided into normal blood glucose group (n=83) and elevated blood glucose group (n=67), then aspirin resistance rate was compared between two groups.Logistic regression analysis was used to analyze influencing factors for aspirin resistance.Results: Compared with aspirin sensitive group, there was significant rise in body weight[(63.5±7.9) kg vs.(66.9±10.2) kg]in aspirin resistance group, P=0.041.Compared with normal blood glucose group, there was very significant rise in aspirin resistance rate (14.46% vs.73.13%) in elevated blood glucose group, P=0.008.Multi-factor Logistic regression analysis indicated that body mass index (BMI) and blood glucose level were independent risk factors for aspirin resistance (OR=4.012, 3.012, P=0.016, 0.018).Conclusion: Incidence rate of aspirin resistance is high in women with ischemic stroke.Blood glucose control and body weight status may affect incidence of aspirin resistance.

Objective To investigate treatment effects of levonorgestrel-releasing intrauterine system (LNG-IUS) for adenomyosis with severe dysmenorrhea in a prospective cohort study. Methods From December 2006 to December 2014, patients of symptomatic adenomyosis diagnosed by transvaginal ultrasound in outpatient or inpatient clinics of Peking Union Medical College Hospital were given the treatment of LNG-IUS. Before and after placement of LNG-IUS, all the patients′parameters were recorded prospectively, including symptoms and scores of dysmenorrhea, menstruation scores, biochemical indicators, physical parameters, carrying status of LNG-IUS, menstruation patterns and adverse effects. Changes of scores and patterns of pain during follow-up were analyzed. Results Totally 1 100 women meets inclusion criteria, among which 640 cases (58.18%, 640/1 100) had severe dysmeorrhea, with median follow-up period of 35 months (range 1-60 months), and accumulative carrying rate of 65% at 60 months follow-up. After placement of LNG-IUS, scores of pain and ratio of severe dysmenorrhea had decreased significantly compared with baselines (all P<0.01), the scroes of visual analog scale (VAS) were 8.1 ± 0.9, 5.5 ± 2.4, 4.6 ± 2.4, 3.3 ± 2.2, 2.2 ± 2.1, 2.2 ± 1.8, 1.4 ± 1.6 and 1.3 ± 1.3 at 0, 3, 6, 12, 24, 36, 48 and 60 months respectively. During 36 months after placement of LNG-IUS, scores of pain had improved significantly compared with preceding period (all P<0.01). We found no universal dependent factors predicting improvement of pain, which was neither relevant with simultaneous changes of menstruation patterns nor adverse effects (all P>0.05). Conclusion LNG-IUS is effective for adenomyosis of severe dysmenorrhea. Improvement of pain is independent on patients characters, menstruation patterns or adverse effects.