Objective To investigate the expression and clinical significance of androgen receptor (AR) and embryonic stem cell associated transcripts 4 (NANOG) in breast cancer patients with human epidermal growth factor receptor 2 (HER-2) over-expression, and to analyze its relationship with clinicopathologic features of breast cancer. Methods 143 breast cancer patients with HER-2 over-expression were selected from the screening of 1052 cases of invasive breast cancer according to estrogen receptor (ER), progesterone receptor (PR) and HER-2 status. The protein expression of AR and NANOG was assayed by using immunohistochemistry.The relationship between AR expression and clinicopathological features was analyzed by χ2 test. The correlation between AR expression and NANOG expression was analyzed by Spearman correlation analysis. Results The positive expression of AR was 35.7 % (51/143). The AR expression was not associated with age and menstruation status (both P>0.05), and was associated with tumor size, clinical TNM staging and lymphatic metastasis (all P< 0.05). The positive rate of NANOG were 53.1 %(76/143), and NANOG proteins were negative in adjacent normal breast tissue and benign breast lesions. The positive rate of AR was 27.6%(21/76) in NANOG-positive cases, whereas the positive rate of AR was 44.8%(30/67) in NANOG-negative cases, and the difference was statistically significant (χ2=4.526, P=0.033). There was an inverse correlation between NANOG and AR expressions (r= -0.255, P= 0.002). Conclusion AR and NANOG may be new targets for endocrine therapy and molecular biological therapy.

Objective To examine the relationship between the concentration of 25-hydroxy vitamin D3 [25-(OH) D3] in the serum and the body mass,the severity of obesity,body mass index(BMI),blood lipid,and their predicting role in obesity children.Methods The study recruited 244 subjects,who see the doctor in Wuxi Maternal and Child Health Hospital,Childhood Nutrition Outpatient from Jul.2011 to Feb.2013.The intake dose of vitamin D each day was investigated,and weight,height,BMI,concentration of 25-(OH) D3 in serum,and microelement were also measured.In addition,lipid metabolism of 38 cases with obesity over 3 years old was determined.Results 1.The serum 25-(OH) D3 concentration of obese children was (68.31 ± 23.06) nmol/L.The concentration of 25-(OH) D3 was lowest in the group of obese children over 36 months of age[(55.03 ± 15.18) nmol/L].2.The concentration of 25-(OH) D3 in the group of obese and overweight children was far lower than that of the children in the normal group (F =4.739,P ＜0.05).3.The concentrations of 25-(OH) D3 in the severely obese children was significantly lower than that of the mild and moderate obesity children(F =9.711,P ＜ 0.05).4.There were significantly inverse associations of serum 25-(OH) D3 with weight,weight and height percentage,BMI (r =-0.365,-0.237,-0.175,all P ＜ 0.001).5.There were significantly inverse associations between the concentration of 25-(OH) D3 in serum with weight,triglyceride in obese children more than 3 years old (r =0.476,-0.324,all P ＜ 0.05).Conclusions The decreasing level of 25-hydroxy vitamin D3 in the serum was associated with obesity.The cause of it might be the increase of the obese adipose tissue,vitamin D getting trapped in fat cells,and all these factors can lead to a less serum vitamin D levels.The vitamin D consumption of obese children is higher than that of normal children,and should supply more vitamin D to reach normal 25-(OH) D3 level.

Objective To analyze the risk factors of colorectal adenoma,construct a nomogram prediction model for the risk of colo-rectal adenoma,and evaluate its predictive efficiency.Methods A total of 1066 patients who underwent colonoscopy in Qingdao Munici-pal Hospital from January 2018 to June 2019 were collected retrospectively.Among them,496 patients who were pathologically diagnosed with colorectal adenoma were included in the adenoma group,the other 570 patients had no obvious abnormality in colonoscopy,or patho-logical diagnosis of inflammatory or hyperplastic polyps were included in the non-adenoma group.The independent risk factors for color-ectal adenoma were analyzed by univariate and multivariate analysis;R software was used to establish a nomogram model for predicting the risk of colorectal adenoma,and the Bootstrap method was used for internal validation of the model.The Calibration curve and receiver op-erator characteristic curve was used to evaluate the predictive efficiency of the nomogram.Results The results of univariate and multiva-riate analysis showed that advanced age(OR=1.068,95％CI:1.049-1.087,P＜0.001),male(OR=0.593,95％CI:0.396-0.886,P=0.011),alcoholism(OR=1.847,95％CI:1.085-3.144,P=0.024),family history of tumor(OR=1.778,95％CI:1.241-2.547,P=0.002),changes in bowel habit(OR=3.508,95％CI:2.496-4.930,P＜0.001),diabetes(OR=1.832,95％CI:1.179-2.846,P=0.007),high neutrophil/lymphocyte ratio(OR=2.861,95％CI:2.055-3.982,P＜0.001),high carcino-embryonic antigen(OR=1.391,95％CI:1.229-1.574,P＜0.001)were independent risk factor for colorectal adenoma.The C-in-dex of the nomogram prediction model was 0.780,and the C-index of the validation model was 0.774.Conclusion The nomogram pre-diction model constructed in this study has high prediction accuracy,which can provide a basis for screening high-risk groups of colorec-tal adenoma and provide a reference for clinicians before endoscopy.

Polycystic ovary syndrome(PCOS)is a heterogeneous disorder closely associated with reproductive endocrine dysfunction in the women.The etiology and pathogenesis of PCOS remain unclear.PCOS is the result of the combination of endocrine metabolic disorders,genetics,and environmental factors.Hyperandrogenemia(HA)and insulin resistance(IR)are the fundamental pathophysiological changes in the development of PCOS,and their interactions exacerbate the clinical manifestations of the PCOS patients.The family aggregation and twin study results confirm the genetic predisposition of PCOS;the genome-wide association study(GWAS)results confirm some risk loci and candidate genes of PCOS.The unhealthy lifestyle habits and environmental endocrine disruptors also play an important role in the progression of PCOS,and the gut microbita is involved in the pathogenesis of PCOS.This article provides a comprehensively retrospective analysis on the recent studies about PCOS,and reviews both internal factors and external factors related to the etiology and pathogenesis of PCOS.

Objective:To develop an automated landmark location system applicable to the case of landmark missing.Methods:Four and eighty-one lateral cephalograms, which contained 240 males and 241 females, with an average age of (24.5±5.6) years, taken from January 2015 to January 2021 in the Department of Orthodontics, Capital Medical University School of Stomatology, and met the inclusion criteria were collected. Five postgraduate orthodontic students were the annotators to manually locate 61 possible landmarks in 481 lateral cephalograms. Two assistant professors in the department as reviewers performed calibration. Two professors as arbitrators, made final decision. Data sets were established (341 were used as training set, 40 as validation set, and 100 as test set). In this paper, an automatic landmarks identification and location model based on convolutional neural networks (CNN), CephaNET, was developed. The model was trained by feeding the original image into the feature extraction module and convolutional pose machine (CPM) module to locate landmarks with high accuracy using deep supervision. Training set was enhanced to 1 684 images by histogram equalization, cropping, and adjustment of brightness. The model was trained to compare the Gaussian heat maps output from the network with the set threshold to identify landmark missing cases. Test set of 100 lateral cephalograms was used to test the accuracy of the model. The evaluation criteria used were success detection rate of missing landmark, mean radial error (MRE) and success detection rate (SDR) in the range of 2.0, 2.5, 3.0, 3.5 and 4.0 mm.Results:The model identified and located 61 commonly used landmarks in 0.13 seconds on average. It had an average accuracy of 93.5% in identifying missing landmarks. The MRE of our testing set was (1.19±0.91) mm. SDR of 2.0, 2.5, 3.0, 3.5 and 4.0 mm were 85.4%, 90.2%, 93.5%, 95.4%, 97.0% respectively.Conclusions:The model proposed in this paper could adapt to the absence of landmark in lateral cephalograms and locate 61 commonly used landmarks with high accuracy to meet the requirements of different cephalometric analysis methods.

OBJECTIVETo investigate the changes and roles of follicular regulatory T cells (Tfr) and follicular T helper cells (Tfh) in the pathogenesis of children immune thrombocytopenia (ITP).

METHODS32 untreated ITP patients, as well as 20 healthy controls were enrolled in this study. The proportion of circulating Tfr and Tfh cells were determined by flow cytometry; real-time PCR was performed to detect the expression of transcription factors and regulatory factors of Bcl-6, c-Maf, Blimp-1 and PD-1 mRNA; ELISA was used to detect plasma concentration of IL-2, IL-6, IL-10 and IL-21.

RESULTS(1)The proportion of Tfh cells were significantly higher (P<0.05), while the Tfr cells and the ratio of tfr/Tfh cells in ITP patients were significantly lower than that in health controls (P<0.05); (2)Correlation analysis showed that the Tfr cells and the ratio of Tfr/Tfh were positively correlated with the platelet counts and negatively with the levels of PA-IgG, while the proportion of Tfh cells was positively correlated with the levels of PA-IgG and negatively with the platelet counts in peripheral blood; (3)Transcription levels of Bcl-6 and c-Maf mRNA in CD4(+) T lymphocytes cells were significantly elevated, the Blimp-1 mRNA in CD4(+) cells and PD-1 mRNA levels of Treg were lower in ITP patients in comparison with healthy controls; (4)The higher Plasma concentration of IL-21, and lower concentration of IL-2 were found in ITP patients.

CONCLUSION(1)The lower proportion of Tfr cells and higher proportion of Tfh cells, as well as the abnormal ratio of Tfr/Tfh might account for the decreased platelet counts to be further involved in the immunological pathogenesis of children ITP; (2)The changes of plasma cytokines IL-2, IL-21 in microenvironment and the over-expression of Bcl-6 mRNA, c-Maf mRNA and the lower-expression of Blimp-1 mRNA in CD4(+) T cells, and over-expression of PD-1 mRNA in Treg cells might be account for the abnormal ratios of Tfr/Tfh cells in ITP patients.

Objective To clarify the effect of iodine intake on serum thyroglobulin (Tg). Methods A 5-year prospective study was conducted in the 3 different iodine intake areas in China [Panshan (miht deficiency) ,Zhangwu (more than adequate) and Huanghua (excess)]. A total of 3 099 people with normal serum levels of Tg in 1999 were followed and 2 448 of these participants were feasible to be observed in 2004 and included in the present study. The serum levels of Tg, thyraglobulin antibody(TgAb), thyroid peroxidase antibody(TPOAb) and TSH, thyroid volume, family and personal histories of thyroid diseases were measured and inquried. The general linear model (GLM) was used to explore the determinants of Tg. Results Among the study population at baseline, serum Tg were significantly different in three areas [7.5 (4.4-13. 1) μg/L at Panshan, 6.8 (3.6-11.2)μg/L at Huanghua, 5.9 (3.2-10.7) μg/L at Zhangwu, P<0.01]. They were associated with age, sex and the rate of positive TgAb, abnormal thyroid volume, abnormal TSH and positive personal history of thyroid diseases, in order to control the effects of confounding factors, the data from 1856 subjects with thyroid-related indexes all in normal range and without personal history of thyroid diseases were analyzed to clarify the effect of iodine intake on Tg. The serum Tg among three areas were significantly different in both 1999 and 2004, they were all increased in 5 years with significant augment (△ Tg) among the three areas[3.1 (-0.2-8.0) μg/L at Panshan, 3.5 (0.5-9.0)μg/L at Huanghua vs 2. 5(0.3-6.1) μg/L at Zhangwu,P<0.01]. The GLM analysis revealed that age, Tg and TSH levels at baseline were the determinants of △Tg in addition to iodine intake. Conclusion Iodine intake is a dominant determinant of serum Tg. Age and TSH should also be considered while indicating iodine intake by serum Tg.

Based on the rDNA sequence of Pichia pastoris, a multi-copy gene expression vector of transglutaminase (pPICZα-rDNA-mtg) was constructed and transformed to the host strain (pGAP9-pro/GS115) expressing pro peptide, to obtain the co-expression strain pro/rDNA-mtg (GS115). Real-time fluorescence quantitative PCR (qPCR) was used to analyze transglutaminase gene copy number in the 4 positive strains. We further studied the effect of gene copy on the enzyme production of recombinant Pichia pastoris as well as high-density fermentation of higher expression strain in a 3-L fermenter. The mtg copy numbers of the 4 positive strains were 2.21, 3.36, 5.72 and 7.62 (mtg-2c, mtg-3c, mtg-6c and mtg-8c), respectively, and the enzyme production capacity and protein expression level were mtg-3c>mtg-2c>mtg-6c>mtg-8c. Mtg-3c and mtg-6c of high-density fermentation had the highest enzymatic activity and enzymatic activity per unit wet weight in the supernatant of 3.12 U/mL, 52.1 U/g (wet weight) and 2.07 U/mL and 36.5 U/g (wet weight), respectively. In terms of enzyme activity per unit wet weight, mtg-3c is 1.4 times higher than that of mtg-6c. The activity of purified enzyme (mtg-3c) was up to 7.21 U/mL and the protein concentration was 437.2 μg/mL. By analyzing the effect of mtg copy number on the enzyme production of recombinant strains, mtg-3c is suitable for the co-expression of two genes (pro and mtg) in pro/rDNA-mtg, and its enzyme activity is related to higher protein secretion of the strain.

Objective To describe a rhesus monkey model of temporal lobe epilepsy (TLE) established via repetitive unilateral intra-amygdala kainic acid (KA) injection and provide experimental basis for epileptogenic network and related research. Methods Eight male adult rhesus monkeys were randomly divided into KA injection group (n=6) and saline injection group (n=2). Brain stereotaxic technique, micro catheter implantation into the right amygdaloid nucleus, subcutaneous bladder connection, and continuous video-EEG monitoring were performed, and KA or saline injection into their right amygdala was achieved. Interictal epileptic discharges (IEDs), ictal discharges and behavioural performance between the two groups were compared right after injection and within 6 months of first discharge. Results Typical IEDs were recorded in the 6 monkeys from KA injection group after 2-4 times of KA injection, with focal spike waves discharges at the right temple area as manifestation; ictal discharges were recorded in 4 monkeys, with discharge patterns of discharges from the right temple area to the whole brain as manifestation, and during epileptic attack, these 4 monkeys suddenly stopped and dumbfounded without obvious limb seizures. Monkeys from the saline injection group showed no obvious abnormal behaviors. Conclusion Through a modified protocol of unilateral repetitive intra-amygdala KA injection, a rhesus monkey model with high similarity of behavioral and brain electrical features to TLE is developed.

OBJECTIVE：To analyze th e general characteristics and application of the models used in the pharmacoeconomic evaluation of type 2 diabetes mellitus （T2DM）therapy during the past 5 years，and to provide reference for the selection and improvement of T 2DM pharmacoeconomic model. METHODS ：Retrieved from PubMed ，Embase，CNKI，Wanfang database and VIP during Jan. 1st，2015 to Dec. 31st，2019，pharmacoeconomic evaluation literatures about T 2DM therapy were collected ；the included model was analyzed in respects of general structure ，therapy plan establishment ， short-term therapeutic efficacy ， complication simulation ，model effectiveness validation ，application frequency. RESULTS ：A total of 81 literatures were included ， involving 14 models，such as CORE model ，Cardiff model ，ECHO model ，etc. Mostly ，Markov or micro Markov simulation method were adopted to measure the patient ’s lifetime health outcome and cost mostly from the point of view of third-party payer. Seven models could simulate 2-4 therapy plans ；short-term efficacy mainly included risk factors of diabetic complications （such as glycosylated hemoglobin level and body mass ）and adverse drug reactions. Most models used intermediate indexes to simulate the occurrence of complications ，and the number of complications ranged from 3 to 15；the validity of model effectiveness included surface validity ，internal validity and external validity ，etc. Among 14 models，the most frequently used models in the past 5 years were long-term models that had been validated ，among which CORE model had the most application times （38/81，46.91％）， followed by Cardiff model （12/81，14.81％）. CONCLUSIONS ：The 14 models have similar structure. The differences of the models are mainly reflected in 3 aspects as therapy plan setting， considered short-term efficacy ， the number of model are ideally choose based on available evidences.