ObjectiveTo investigate the association between serum creatinine/cystatin C ratio （CCR） and nonalcoholic fatty liver disease （NAFLD） based on the NHANES database， and to evaluate the potential significance of CCR as an indicator reflecting the metabolic status of the body. MethodsBased on the data from the NHANES database in 1999‍ ‍—‍ ‍2004， a total of 4 217 participants were enrolled and divided into NAFLD group with 1 726 participants and non-NAFLD group with 2 491 participants. CCR was compared between the two groups， and the association between CCR and NAFLD was analyzed. The Wilcoxon rank-sum test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups. The multivariate logistic regression model was used to investigate the association between CCR and NAFLD； CCR was divided into 4 groups based on quartiles， and odds ratio （OR） and 95% confidence interval （CI） in the regression model was calculated with the first quartile as reference. In addition， the restricted cubic spline analysis was used to investigate whether there was a non-linear relationship between CCR and NAFLD， and interaction items were introduced into the Logistic regression model to perform an interaction analysis. Subgroup analyses were performed based on the stratification of variables to investigate the difference in the association between CCR and NAFLD in different populations. ResultsThe non-NAFLD group had a significantly higher CCR than the NAFLD group （Z=-4.76，P<0.01）. The Logistic regression analysis showed that in model 1 without adjustment of variables， CCR was negatively associated with NAFLD （OR=0.993，95%CI：0.989‍ ‍—‍ ‍0.996，P<0.01）， and in model 3 with adjustment of all variables， CCR was still negatively associated with NAFLD （OR=0.986，95%CI：0.981‍ ‍—‍ ‍0.991，P<0.01）. The analysis of CCR based on quartiles showed a significant association between the increase in CCR and the reduction in the risk of NAFLD. In model 3， compared with the individuals with the lowest quartile of CCR， the individuals with the highest quartile of CCR had a significantly lower risk of NAFLD （OR=0.426，95%CI：0.316‍ ‍—‍ ‍0.574，P<0.01）. Further interaction and subgroup analyses showed that the interaction between CCR and age/sex had a statistical significance （P_interaction<0.01 and P_interaction=0.04）. The subgroup analysis based on age showed a more significant association between CCR and NAFLD in the middle-aged population （≤60 years） （OR=0.982，95%CI：0.976‍ ‍—‍ ‍0.987）， and the subgroup analysis based on sex showed a stronger association between CCR and NAFLD in women （OR=0.979，95%CI：0.972‍ ‍—‍ ‍0.986）. ConclusionThis study shows a significant negative association between CCR and NAFLD， and such association is more significant in middle-aged individuals and women.

Epigenetic mechanisms play a crucial role in the development and progression of nonalcoholic fatty liver disease （NAFLD）， especially among lean individuals. The research on related epigenetic mechanisms has provided new clues and directions for revealing the underlying causes and treatment strategies of NAFLD. This article introduces the role of epigenetics in the development and progression of NAFLD among lean individuals in recent years， analyzes the latest research advances in the epigenetics of NAFLD in this population， and briefly describes the basic concepts of epigenetics， including DNA methylation， histone modifications， and non-coding RNA regulation. This article also discusses how epigenetic alterations impact the pathogenesis， disease progression， and treatment strategies of NAFLD in lean individuals.

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 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.

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.

Acetaminophen (APAP) overdose is the leading cause of drug-induced liver injury, and its prognosis depends on the balance between hepatocyte death and regeneration. Sirtuin 6 (SIRT6) has been reported to protect against oxidative stress-associated DNA damage. But whether SIRT6 regulates APAP-induced hepatotoxicity remains unclear. In this study, the protein expression of nuclear and total SIRT6 was up-regulated in mice liver at 6 and 48 h following APAP treatment, respectively.

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.

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 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 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.