Objective: To explore the influence factors of the degenerate oligonucleotide primered PCR(DOP-PCR). Methods: Genome DNA template from the mouse single oocyte or liver tissue were used to perform DOP-PCR. DOP-PCR was carried out with templates of different origin, different gradient dilution, with or without low melting point gel purified to wipe off the small fragment that might interfere with the following analysis, and then PCR of gene FTCD and CBS were carried out to evaluate the influence of these factors on the amplification efficiency and specificity. Results: Compared with genome DNA template from mouse liver, the template from single oocyte had the same efficiency and specificity but a minor yield and different gradient dilution of DNA template had no effect on the efficiency and specificity. Furthermore, there was a higher specificity in the low melting point gel-purified DOP-PCR product than in untreated ones. Conclusion: We have got a satisfactory result and increased specificity from DOP-PCR product purified with the low melting point gel. Single oocyte of mice could be used for further investigation of special genes detection by DOP-PCR and of an optimization in the yield of the products.

Objective: To study the correlation between serum resistin (SR) level and blood pressure in patients with type 2 diabetes mellitus (T2DM) complicated hypertension. Methods: A total of 124 patients with T2DM complicated hypertension (T2DM + hypertension group), 85 patients with pure T2DM (pure DM group) and 71 normal subjects (normal control group) were enrolled. According to blood pressure level, T2DM + hypertension group was further divided into stage 1 subgroup (n=34), stage 2 subgroup (n=43) and stage 3 subgroup (n=47). Peripheral venous blood was taken to measure levels of SR and other indexes in three groups. Logistic multi-factor regression analysis was used to analyze the risk factors affecting blood pressure level. Kendall’s tau-b correlation analysis was used to analyze the correlation among SR, fasting insulin, quantitative insulin sensitivity check index, homeostasis model-insulin resistance index and blood pressure level. Results: Compared with normal control group, there were significant rise in SR levels [(7.21±2.65) ng/ml vs. (19.87±3.67) ng/ml vs. (31.32±3.89) ng/ml] in pure DM group and T2DM + hypertension group, and that of T2DM + hypertension group was significantly higher than that of pure DM group, P<0.01 all. Logistic multi-factor regression analysis indicated that SR level was an independent risk factor influencing blood pressure level of patients (R2=0.087, P<0.05). Kendall’ s tau-b correlation analysis indicated that SR level was significantly positively correlated with blood pressure level of patients with hypertension (r=0.202, P=0.001); after other influencing factors were controlled, the significant positive correlation still existed between them (r=0.233, P=0.022). Conclusion: Serum resistin level rise is a risk factor of patients with type 2 diabetes mellitus complicated hypertension, and it is significantly positively correlated with their blood pressure level.

Objective To evaluate the effect of the traditional Chinese medicine (TCM) knowledge and skills training on non TCM community physicians in Shanghai, and provide important basis and feedback for future adjustments and training improvement.Methods The current situation of the TCM knowledge and skills training on non TCM community physicians in Shanghai was reviewed. The community physicians participated in the training courses in 2014 in Jiading district were randomly selected for investigation. The TCM theory and skills scores of all the 65 trainees were compared before and after training, a self-made questionnaire survey on assessment of training was conducted after training, and the influencing factors of training effect were analyzed. Results After training,test results of TCM theory and skills were significantly higher than before(P<0.01). According to the survey of trainees after TCM training, 84.6% thought the training was necessary,95.4% liked the teaching form, 93.9% accredited training content difficulty level,92.3% thought the training was practical, 98.5% were willing to provide Chinese medicine services for community residents, 60.0% thought training to promote the general medical work was very helpful. The existing problems were short training periods(43.1%), only 10.8% could skilled TCM appropriate technologies and 60.0% were basically master. Conclusion The TCM knowledge and skills training on non TCM community physicians in Shanghai had achieved better effect. There still were some deficiencies, which require further exploration in order to better carry out future training.

Objective: To clone a novel gene and explore its expression patterns in tissues and cells,so as to find its role in the process of encephalopathy in DS.Methods: On the base of our previous microarray's result together with the tissue type,we chose EST AI480014 to carry out RACE,then analyzed its expression profiles in liver,spleen,kidney,heart,brain by multi-tissues Northern blot,after that semi-quantitive RT-PCR was used to reexamine the expression profiles.Furthermore,we used ISH to find whether aim gene expressed in neuroglial cells cultured in vitro.Finally we performed semi-quantitive RT-PCR to explore whether it expressed differently between DS and normal.Results: We gained a 682 bp new cDNA fragment(DQ275636)which expressed in all the tissues examined and had no alternative splices in them.It expressed highly in brain especially in frontal lobe and hippocampus.According to the ISH result we convinced that it expressed in neuroglial cells.Using bioinformatics we mapped DQ275636 to chromosome 5q14.Conclusion: We have obtained a new gene fragment based on the(above) results.According to its expression character and tissue type,it can be suggested that this gene has a probable role in the process of encephalopathy in DS.

Objective:To discuss the causes and effective measures of prevention and treatment of chylous fistula after central lymph node dissection (CLND) of thyroid cancer. Methods:A total of 6 127 patients who underwent CLND of thyroid cancer in the Tianjin Medical University Cancer Institute and Hospital between July 2013 and June 2015 were analyzed;of which, 14 patients acquired the complication of postoperative chylous fistula. The following conservative treatments were initially performed:systemic therapy, local pressure bandaging, normal pressure drainage, 50%glucose injection, or pingyangmycin injection through a drainage tube. Surgical op-eration was then conducted when the efficacy of the treatment was poor. Results:After the conservative treatment of the 14 patients, the drainage volume gradually decreased in 12 patients, and surgery was performed on the remaining two patients. Conclusion:The CLND of thyroid cancer must be carefully conducted to prevent postoperative chylous fistula. An active conservative treatment must be the first option when chylous fistula occurs. Surgery must only be performed if the treatment is invalid.

BACKGROUNDAbnormal neuronal differentiation plays an important role in central nervous system (CNS) development abnormalities such as Down syndrome (DS), a disorder that results directly from overexpression of genes in trisomic cells. Receptor-interacting protein 140 (RIP140) is significantly upregulated in DS brains, suggesting its involvement in DS CNS development abnormalities. However, the role of RIP140 in neuronal differentiation is still not clear. The current study aimed to investigate the effect of RIP140 overexpression on the differentiation of neuro-2a (N2a) neuroblastoma cells, in vitro.

METHODSStably RIP140-overexpressing N2a (N2a-RIP140) cells were used as a neurodevelopmental model, and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot. Retinoic acid (RA) was used to stimulate N2a differentiation. Combining the expression of Tuj1 at the mRNA and protein levels, the percentage of cells baring neurites, and the number of neurites per cell body was semi-quantified to determine the effect of RIP140 on differentiation of N2a cells. Furthermore, western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP140 induces differentiation of N2a cells. Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.

RESULTSCompared to untransfected N2a cells RIPl40 expression in N2a-RIP140 cells was remarkably upregulated at both the mRNA and protein levels. N2a-RIP140 cells had a significantly increased percentage of cells baring neurites, and numbers of neurites per cell, as compared to N2a cells, in the absence and presence of RA (P < 0.05). In addition, Tuj1, a neuronal biomarker, was strongly upregulated in N2a-RIP140 cells (P < 0.05) and phosphorylated ERK1/2 (p-ERK1/2) levels in N2a-RIP140 cells were dramatically increased, while differentiation was inhibited by the ERK1/2-specific inhibitor U0126.

CONCLUSIONSRIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.

Blotting, Western ; Cell Differentiation ; physiology ; Cell Line ; Humans ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Neurons ; cytology ; metabolism ; Nuclear Receptor Co-Repressor 1 ; metabolism ; Signal Transduction ; physiology

Objective To study the differences of glycoprotein gene (G gene) between rabies virus epidemic strains of Guizhou province in recent years and vaccine strains,and to provide scientific basis for the development of rabies vaccine and establishment of effective control and prevention measures.Methods RT-PCR assay was used to amplify the G gene of rabies positive brain tissues samples of human and dog derived from Guizhou province in recent years.The amplification products were sequenced and comparatively analyzed with that of vaccine strains by using bioinformatics software.Results Eight full-length G gene sequences were obtained by RT-PCR amplification,sequencing and splicing.The homogeny of G gene between 8 epidemic strains of Guizhou province and 9 vaccine strains were 82.0％-94.1％ and 87.6％-97.5％ on nucleotide and deduced amino acid level,respectively,and the highest homogeny were found with the human vaccine strain CTN (87.0％-94.1％ for nucleotide and 93.7.％-97.5％ for amino acid) among the 6 human rabies vaccine strains,while highest homogeny were found with strain Flury (83.9％-84.6％ for nucleotide and 91.1％-93.0％ for amino acid) among the three animal vaccine strains.Besides,among the 8 epidemic strains from Guizhou province,strain GZ09 collected in the year of 2005 was of the highest homogeny with human rabies vaccine strain CTN and animal rabies vaccine strain Flury,while strain GZ30 collected in the year of 2010 was of lowest homogeny with human rabies vaccine strain CTN and animal rabies vaccine strain Flury.Moreover,phylogenetic analysis based on the G gene indicated that the relationship of 8 epidemic strains derived from Guizhou,the 9 vaccine strains and genotype 1 Lyssavirus were clustered to a same branch.Vaccine strain CTN among the 9 vaccine strains was closest to the 8 epidemic strains,and the other 8 vaccine strains were relatively more distant from the epidemic strains of Guizhou province.In addition,phylogenetic analysis indicated that among the 8 epidemic strains from Guizhou province,strain GZ09 collectcd in the year of 2005 was of closest evolutionary relationship to CTN,while the other 7 epidemic strains were relatively more distant from CTN.Conclusion This study confirmed on the G gene level that rabies virus strains circulated in Guizhou province in recent years and the vaccine strains used in China belonged to rabies virus genotype 1,and the virus strains circulated in Guizhou province in recent years is of smallest difference with the human vaccine strain CTN and animal vaccine strain Flury.Besides,as time goes on,the difference between the epidemic strain and the vaccine strains becomes more and more obvious.The results of this study will provide scientific basis for the development of rabies vaccine and establishment of effective control and prevention measures.

Objective:To identify Down syndrome (DS) fetal encephalopathy related genes and signaling pathways via bioinformatics analysis, and to explore their potential mechanisms underlying the occurrence and development of DS neuropathology.Methods:Retrospective study.In December 2021, dataset GSE59630 was downloaded from the gene expression omnibus (GEO), and differentially expressed genes (DEGs) between DS and normal fetal brain tissue were identified by R software.Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and gene set enrichment analysis (GSEA) were performed on the genes identified.The protein-protein interaction (PPI) network was constructed based on search tool for the retrieval of interacting genes online database and Cytoscape software, and key modules and hub DEGs were identified.Real-time quantitative polymerase chain reaction technique was used to verify the expression of hub genes related to neurodegeneration in brain tissue of 3 pairs of DS and normal fetuses at the gestational age of 22-33 weeks.Results:A total of 225 DEGs were screened out from DS and normal fetal brain tissue, including 18 up-regulated genes and 207 down-regulated genes.GO functional enrichment analysis showed that DEGs were mainly enriched in neurogenesis, neuronal apoptosis, transcriptional regulation, mitochondrial energy metabolism, etc.KEGG pathway enrichment analysis revealed that DEGs were associated with a variety of neurodegenerative diseases.GSEA suggested that apoptosis and inflammatory responses play a vital part in the occurrence of DS neuropathology.Ten hub genes were identified by the PPI network established, and they were mainly related to histone acetylation and transcriptional regulation.According to the tissue verification result, the variations of RAB8A, TBP and TAF6 expression conformed to the microarray data. Conclusions:The key genes and signaling pathways identified by transcriptome analysis of fetal brain tissue facilitate the comprehensive understanding of the molecular mechanism of DS neuropathology.This study provides a novel insight into the clinical diagnosis and treatment of neurodevelopmental abnormalities and mental retardation in DS.

Palliative care is the final link in the whole lifecycle health management and is an important part of promoting a healthy China. In order to further improve service efficiency and the quality of end-of-life care, Shanghai Jiading Yingyuan Hospital has constructed a new interdisciplinary team collaboration model for palliative care from the perspective of integrated medical theory, and applied this model to clinical practice. By elaborating on the theory of team integration, member composition and division of labor, implementation process and preliminary effects, this study aims to provide a theoretical basis and reference for other regions to carry out the integration path and collaborative model of interdisciplinary services in palliative care.

Objective:To evaluate the effect of dual-tube epidural segmental injection of lidocaine analgesia on the delivery outcome and maternal and infant complications of persistent posterior occipital position postpartum or lateral occipital position postpartum patients with protracted active phase.Methods:The full and single-term primiparas ( n=216, 37 to 42 weeks gestation, 22 to 35 years) diagnosed as persistent posterior or lateral occipital position during the active period were selected from the Department of Obstetrics of Qingdao Municipal Hospital from January 2015 to October 2019. The subjects were randomly assigned into two groups: double-tube epidural block group ( n=108) and single-tube epidural block group ( n=108), 1% lidocaine was used for epidural analgesia respectively under ultrasound guidance. Senior midwife or obstetricians implement new partogram, and guide women to perform position management, and push or rotate the fetal head in a timely manner. Observation indicators: general condition, the use of non-pharmacological analgesic measures, analgesia related conditions and pain visual analogue scale (VAS) score, delivery-related indicator, cesarean section indication, anesthesia-related indicator, maternal and child complications. Results:(1) General condition: the age, weight, height, gestational age, the ratio of persistent lateral or posterior occipital position, cephalic score, and neonatal birth weight between the two groups of women were not statistically significant (all P>0.05). (2) The use of non-pharmacological analgesic measures: the women’s Lamaze breathing method, Doula delivery companionship, percutaneous electrical stimulation, and other measures between two groups were compared, and there were not significant differences (all P>0.05). (3) Analgesia related conditions and VAS scores of women undergoing vaginal delivery: compared with the single-tube epidural block group ( n=40), the second-partum time of the women in the double-tube epidural block group ( n=59) was significantly shortened [(124±44) vs (86±33) minutes, P<0.01]; after 30 minutes of analgesia (4.4±0.5 vs 0.9±0.5, P<0.01), during forced labor in the second stage of labor (5.7±0.6 vs 1.3±0.4, P<0.01), the VAS scores of pain were also significantly reduced ( P<0.01). (4) Labor-related indicators: compared with the single-tube epidural block group, the natural delivery rate (21.3% vs 49.1%) and the delivery experience satisfaction rate (51.9% vs 98.1%) of women in the double-tube epidural block group were significantly increased (all P<0.01), cesarean section rate (63.0% vs 45.4%), instrument assisted rate (15.7% vs 5.6%) decreased significantly (all P<0.05). (5) Cesarean section indications: compared with the single-tube epidural block group, the cesarean section rate caused by prolonged labor or protracted active phase of women in the double-tube epidural block group was significantly reduced (38.0% vs 22.2%； P<0.05), and the fetal distress, intrauterine infection, and social factors caused by cesarean section between the two groups were compared, while the differences were not statistically significant (all P>0.05).(6) Anesthesia related indexes: the block planes of the maternal upper tube administration in the double-tube epidural block group were mostly T7, T8, T9-L2 and L3,While,the block planes in the single-tube epidural block group were mostly T10, T11-S1, S2, S3, and the modified Bromage score were all 0. (7) Maternal and child complications: compared with the single-tube epidural block group, the postpartum hemorrhage rate (18.5% vs 7.4%), the perineal lateral cut rate (20.4% vs 5.6%), the neonatal asphyxia rate (12.0% vs 3.7%), ICU rate of transferred neonates (13.9% vs 4.6%) in the double-tube epidural block group were significantly reduced (all P<0.05). Soft birth canal injury rate, puerperal disease rate and neonatal birth rate between two groups were compared, and there were not statistically significant differences (all P>0.05). Conclusion:Dual-tube epidural segmental injection of lidocaine analgesia could increase the natural delivery rate of women with posterior occipital or lateral occipital position with active stagnation, reduce the rate of cesarean section and the rate of transvaginal instruments, and reduce the complications of mother and child.