ObjectiveTo study the transcript atlas of cell immunity-associated genes in 8 liver cell types including hepatocytes, bile duct epithelial cells, oval cells, hepatic stellate cells, sinus endothelial cells, Kupffer cells, pit cells and dendritic cells from the rat regenerating liver. Methods Isolation of rat 8 liver cell types, detection of their genes expression change and the prediction physiological activities were accorded to cluster program, and the methods of bioinformation and systematic biology, and their gene expression patterns were analyzed by Microsoft Excel software. Results A total of 40 blood coagulation-associated genes yielded the meaningful expression changes in liver regeneration. Serpine1, a2m in eight liver cell types, vwf, klkb1 in seven liver cell types other than Kupffer cells, and other genes in the two or more liver cell types yielded the meaningful expression changes. It suggested that the synthesis of kallikrein and prothrombin、the formation of thrombin were enhanced at the priming and progressing phases of liver regeneration. The biological activities, such as fibrin monomer aggregated into fibrin polymers, were elevated at termination phase.Conclusion The rat liver regeneration is closely associated with the blood coagulation.

Objective To analyze the clinical and molecular genetic characterizations of X-linked adrenal dysplasia congenita(AHC) onset in infant.Methods Seven children (from 7 families) with X-linked AHC who were admitted to the Department of Endocrinology,Genetics and Metabolism,Children's Hospital Affiliated to Zhengzhou University,from July 2012 to June 2017 were selected.All patients were screened for dosage-sensitive-sex reversal-adrenal hypoplasia congenital critical region on the X chromosome gene1 (DAX1/NR0B1) mutations.The clinical manifestation and laboratory examination were analyzed,their clinical characterizations were summarized.Results Seven patients were all male,the onset age of the patients were from after birth to 7 months old,and 4 patients (4 families) had a family history of X-linked recessive inheritance.The clinical manifestations were skin pigmentation [100.0％ (7/7 cases)],vomiting [71.4％ (5/7 cases)],no weight gain [57.1％ (4/7 cases)] and poor spirit [28.6％ (2/7 cases)].Laboratory tests showed that hyperkalemia and hyponatremia,increased coricotrophin,normal or decreased cortisol,17α-hydroxyprogesterone,progesterone,aldosterone and dehydroepiandrosterone.Testosterone levels increased in 5 patients.The abnormalities of adrenal glands imaging could be seen in 2 patients.Two patients were misdiagnosed as congenital adrenal cortical hyperplasia.Then the definitive diagnosis were made by genetic test.DAX1/ NR0B1 gene mutations were found in all patients.Five patients were novel mutations (c.114_126del,c.872G ＞ A,c.56delG,c.884T ＞ G,c.1217delG).Conclusions The clinical manifestations of X-linked AHC with infant onset include pigmentation,poor spirit and growth retardation,which should be differentiated from congenital adrenal cortical hyperplasia.Hormone levels such as elevated blood 17α-hydroxyprogesterone and family history are the main identification points,and AHC cannot be excluded when testosterone level increases.Five novel mutations are found in this study,which enrich the gene database.

Angelica dahurica is diaphoretic, commonly used in the treatment of cold, wet, and itchy rubella nasosinusitis. Studies have shown that the effective components of Angelica dahurica can be used in the treatment of malignant tumors. This paper summarizes the related literature home and abroad recently, and regards the effective components of Angelica dahurica as a major role in treatment of tumor by inhibiting tumor cell proliferation, promoting apoptosis, inhibiting tumor metastasis, inhibiting platelet aggregation, enhancing immunity, and anti-tumor drug resistance. Meanwhile, the paper finds the shortcomings of the present researches, and hopes to provide reference for the future experiment and clinical research.

Objective:To study the effects of Zuogui Pills on the expressions of miR-133b-3p and RhoA in osteoclasts of postmenopausal osteoporosis rats; To discuss its potential mechanism.Methods:SD female rats were randomly divided into normal group, model group, sham-operation group, and Zuogui Pills group using a random number table method, with 6 rats in each group. The model group and Zuogui Pills group were treated with oophorectomy to construct a rat model of osteoporosis. Zuogui Pills group was orally administered with Zuogui Pills decoction at a concentration of 10 g/kg for 12 consecutive weeks. Colorimetric method was used to measure the serum calcium and phosphorus levels of rats, and ELISA method was used to detect ALP levels. Bone density meter was used to measure the bone density of the femurs of rats in each group. The osteoclast of each group were cultured, and the expressions of RANKL and RUNX2 protein were detected by Western blot. MiRNA sequencing and differential expression analysis were performed on bone tissues of rats. Osteoclasts were treated with miR-133b-3p mimic and its negative control. The cell proliferation activity of osteoclasts was detected by cell counting kit-8 (CCK-8). The osteoclast differentiation activity was detected by the tartrate-resistant acid phosphatase staining. The dual-luciferase reporter assay was used to detect the relationship between miR-133b-3p and RhoA. The "rescue" experiment of miR-133b-3p mimic and RhoA co-expression were used to study the molecular regulatory mechanism of Zuogui Pills on osteoclast activity.Results:Compared with the model group, the bone mineral density of Zuogui Pills group significantly increased ( P<0.05, P<0.01), the levels of calcium and phosphorus in serum increased, the level of alkaline phosphatase ALP decreased ( P<0.05), the expression of RANKL protein decreased, and the expression of RUNX2 protein increased. Sequencing results showed that rno-miR-133b-3p was down-regulated in osteoclasts of postmenopausa osteoporosis rats treated with Zuogui Pills with the maximum difference ( P<0.01). Q-PCR results showed that the expression of miR-133b-3p in osteoclasts of Zuogui Pills group was significantly lower than that of the model group. The upregulation of miR-133b-3p could significantly promote the cell proliferation and differentiation of osteoclasts. RhoA overexpression could reverse the excessive proliferation and differentiation of osteoclasts caused by miR-133b-3p overexpression. Conclusions:RhoA is the target gene regulated by miR-133b-3p. Zuogui Pills can inhibit the activity of osteoclasts by regulating miR-133b-3p/RhoA axis, relieving the symptoms of osteoporosis.

BACKGROUND:Endothelial injury is one of the causes of cardiovascular diseases.Human induced pluripotent stem cells are easy to obtain,have strong differentiation ability,and have less exclusiveness,and their endothelial differentiated cells can be used as ideal cells for cardiovascular disease research. OBJECTIVE:To investigate the effect and mechanism of calycosin on endothelial differentiation of human induced pluripotent stem cells and to provide technical support for microvascular regeneration. METHODS:Human induced pluripotent stem cells were divided into control group and calycosin group(1.25,2.5 μg/mL),and growth factors were added to induce single-layer endothelial differentiation.After the induction of differentiation for 8 days,the positive rate of endothelial cell marker CD144 was detected by flow cytometry.Fluorescent expressions of CD144 and CD31 were detected by the immunofluorescence method.Lentivirus RNAi GFP puromycin was used to silence human-induced pluripotent stem cell Piezo1 mRNA followed by endothelial directed differentiation.After 8 days of differentiation,the positive rate of CD144 in differentiated cells was detected by flow cytometry.The mRNA expression levels of CD144,Piezo1 and MEK were detected by qPCR. RESULTS AND CONCLUSION:(1)Compared with the control group,the positive rate of CD144 was significantly increased in the 1.25 and 2.5 μg/mL calycosin groups(P<0.05).The expressions of CD144,Piezo1,and MEK mRNA were increased in the 2.5 μg/mL calycosin group(P<0.05).The fluorescence expressions of CD144(P<0.01)and CD31(P<0.001)were significantly increased in the 2.5 μg/mL calycosin group.(2)Compared with the shNT group,CD144 positive rate and CD144,Piezo1,MEK mRNA expressions were significantly increased in the shNT + calycosin 1.25,2.5 μg/mL groups(P<0.05).Compared with the shPiezo1 group,the positive rate of CD144 and mRNA expressions of CD144,Piezo1 and MEK had no significant changes in the shPiezo1+calycosin 1.25,2.5 μg/mL groups(P>0.05).(3)It is concluded that 2.5 μg/mL calycosin promotes the differentiation of human-induced pluripotent stem cells into endothelial lineages.Calycosin promotes the downstream MEK expression,thereby promoting the endothelial differentiation of human induced pluripotent stem cells by targeting the expression level of Piezo1.

Objective:To explore the research hotspots and effective promotion paths of post market surveillance and supervise of medical consumables with non-active medical devices.Methods:Data mining methods were used to collect related journal literatures and documents from the websites of China regulatory institutions and the China National Knowledge Infrastructure(CNKI),order sub item data of medical device adverse event reports,extract the MeSH element words of literatures and documents,perform bibliometric analysis and visual display.Results:The number of medical devices adverse event reports in China has been increasing year by year,reaching 694 866 in 2022,in the four statistical years from 2019 to 2022,the number of reports on non-active medical devices and IVD reagents also showed a parallel increasing trend,accounting for about 65.00% of the total number of adverse event reports on medical devices in the year.The bibliometric analysis of journal literature shows that research in this field has received varying degrees of participation from regulatory institutions,universities,medical institutions,and enterprises.Regulatory institutions have contributed 46 articles,accounting for 56.79% of the total number of articles,followed by 28 articles from universities.The co-occurrence analysis shows that hot topic is focused in 5 clusters:quality management,risk management,international experiences discussion and adverse event surveillance and re-evaluation and real-world research.China regulatory institutions attach great importance to post market surveillance and supervise,and have issued more than 20 relevant documents since 2006,focusing on specific topics and gradually deepening around safety and effectiveness.Conclusion:The post market surveillance and supervise of medical devices,especially medical consumables based on non-active medical devices,need to be promoted synchronously in three dimensions:regulatory institutions,medical institutions,and enterprises.Universities,research institutes,and industry organizations should work in coordinating to strengthen the collection,identification,and active surveillance of risk signals based on adverse event surveillance,safety evaluation based on risk management,and conducting real-world research,research and develop risk control and corrective and preventive measures.

Objective To explore the effect of oral feeding promotion program, which combined utilization of preterm infant oral motor intervention ( PIOMI) from abroad and the Chinese version of preterm infant oral feeding readiness assessment scale (PIOFRAS), on the performance, process and outcome of oral feeding in preterm infants.Methods A total of 62 infants born before 34 weeks of gestation were divided into control group and experimental group according to the order of admission. Both groups received the same routine care. The infants in the experimental group ( n=30 ) received the assessment of oral feeding readiness by PIOFRAS before feeding, and were given oral massage and non-nutritive sucking for 5 minutes once a day. The intervention started from gavage feeding to total oral feeding. While the control group ( n=32) received a sham intervention that nurses stood beside the warm box or the baby′s bed for 5 minutes once a day before feeding. The oral feeding performance, the corrective gestational age of initiation of oral feeding and achieving full oral feeding, daily bodyweight and length of hospital stay were recorded. Results The transitional time from the initiation of oral feeding to full oral feeding in experimental group was shorter than that in control group[(8.07± 6.73) d vs (13. 88±7.51)d], and the difference was statistically significant (t=3.200,P<0.05). There were no significant differences in the feeding performance, weight gain and length of hospital stay between two groups (P>0.05).Conclusions The oral feeding promotion program can improve the oral feeding of preterm infants.

Bacillus subtilis can be widely used as an important microorganism for metabolic engineering and recombinant proteins expression in industrial biotechnology and synthetic biology. However, it is difficult to make accurate regulation of exogenous gene by biological tools in B. subtilis, which limits the application of B. subtilis in synthetic biology. The purpose of this study is to develop regulatory tools for precise control of gene expression by using non-coding RNAs, by which the activation of heterologous gene could be achieved without the auxiliary protein factors. We constructed the synthetic riboswitch E and aptazyme AZ using the theophylline aptamer. Six different native promoters from B. subtilis were functionally adapted with the E and AZ to fabricate an array of novel regulatory elements activated by theophylline. Then, we determined the performance of these elements using green fluorescence protein as reporter, and then further verified using red fluorescence protein and pullulanase as cargo proteins. Results showed that the same kind of RNA elements with different promoters showed different levels of efficiency. Promoter PsigW and E combination (sigWE) had the highest induction rate in B. subtilis. Compared with the control group, it can produce the induction rate of 16.8. Promoter PrpoB and AZ combination (rpoBAZ) showed the highest induction rate of 6.2. SigWE mediated mCherry induction rate was 9.2, and P43E mediated pullulanase induction rate was 32.8, in which enzyme activity reached 81 U/mL. This study confirmed that GFP, mCherry and pullulan can all be regulated by riboswitch and aptazyme, but there were differences between different combinations of promoters with RNA regulators.
Bacillus subtilis ; Promoter Regions, Genetic ; RNA ; Recombinant Proteins ; Theophylline

Objective: Understanding mental health status of students with learning disabilities in Beijing to provide a basis for mental health promotion of students with learning disabilities. Methods: By means of random cluster sampling, 5 787 enrolled students in grade one and grade two of 11 public junior middle schools in Beijing were selected as the survey subjects. A self designed questionnaire was used to investigate the students’ learning disabilities and mental health status through anonymous self filling. Results: About 11.6% students self reported learning disabilities. The proportions of students with learning difficulty in mathematical reasoning and calculation were higher, 44.1% and 40.7% respectively. The reported rate of mental health problems was 38.3%. The top four of the 10 symptoms were obsessive compulsive symptoms, learning pressure, emotional instability and anxiety(2.19±0.77)(2.17±0.99)(2.09±0.90)(2.07±1.08). Compared with students without learning disabilities, students with self reported learning disabilities had poorer mental health status(OR=1.47), and learning disabilities were related to most factors of mental health. Different types of learning disabilities were associated with different mental health factors. Conclusion Mental health problems of students with learning disabilities are higher than that of students without learning disabilities. It is necessary to strengthen the mental health support for students with learning disabilities and improve their mental health.

Objective:To understand the genome analysis and molecular typing of foodborne Yersinia enterocolitica ( Y.e) strains in Liaocheng City of Shandong Province from 2020 to 2021. Methods:The Y.e strains were isolated from raw meat and meat products. Then we made the strain identification, drug sensitivity test, virulence gene test, pulsed-field gel electrophoresis (PFGE), and whole genome sequencing (WGS). The genome sequencing data were assembled with the microbial genome annotation package. We performed the multilocus sequence typing (MLST) and core genome multilocus sequence typing (cgMLST) and used WGS-based single nucleotide polymorphism typing (wg-SNPs) method to carry out genetic evolution analysis with 14 domestic and Y.e genomes obtained from the NCBI. Results:A total of 21 strains of Y.e were detected from 165 samples, with a detection rate of 12.73%. The 20 strains of Y.e were sequenced successfully. The 20 strains of Y.e carries a variety of drug resistance genes and virulence genes, showing multiple drug resistance. The virulence gene PCR test showed that 21 strains of Y.e having two virulence genes. Cluster analysis of PFGE, MLST, and cgMLST showed that the genomics of 21 strains was highly diverse. The genetic evolution analysis of wg-SNPs showed that 20 Y.e strains could be divided into two main evolutionary branches. Conclusions:Y.e strains isolated from raw meat in Liaocheng City carry a variety of drug resistance genes and virulence genes, and the molecular typing is highly diverse, which may cause infection risk. The molecular biological monitoring of Y.e in raw meat should be strengthened, and genome sequencing and molecular typing detection be carried out to provide the theoretical basis for foodborne illness caused by Y.e.