ObjectiveUsing the Delphi method and analytic hierarchy process （AHP） to screen diagnostic items for qi stagnation syndrome and determine their weights， providing a reference for the development of a diagnostic scale of qi stagnation syndrome. MethodsLiterature related to qi stagnation syndrome were screened from databases including CNKI， Wanfang， VIP， SinoMed （from inception to October 31， 2020）. Through systematic review of literature and expert discussions， the information on the four examinations of traditional Chinese medicine were organized and an item pool was constructed. The Delphi method was used to screen the item indicators， while the AHP was employed to determine their weights. Statistical methods such as mean value， full score ratio， rank sum， unimportant percentage， and coefficient of variation were used for item screening， with the weights calculated by AHP serving as the item weights. ResultsA total of 235 articles and books were included for analysis， resulting in an item pool of 16 items. After three rounds of expert consultation， a total of 84 valid questionnaires were collected， with a total expert enthusiasm coefficient of 99% and authority coefficient of 0.86， 0.84， 0.83， respectively， and the coordination coefficients were 0.45， 0.49， and 0.29， respectively. Through the statistics analysis， 8 diagnosis items were screened out， including distension （stuffi-ness） or distending pain or scurrying pain， wiry pulse， depressed emotions， frequent sighing， deep and wiry pulse， irritability， pale red tongue， and thin white coating. The AHP showed that the order of weights of the first-level indicators from high to low was clinical symptoms， pulse manifestation， and tongue manifestation； the order of weights of the second-level indicators from high to low was distension （stuffiness） or distending pain or scurrying pain， wiry pulse， depressed emotions， frequent sighing， deep and wiry pulse， irritability， pale red tongue， and thin white coating. ConclusionBy applying the Delphi method and AHP to analyze and evaluate the diagnostic items for qi stagnation syndrome， key diagnostic items were screened and their weights determined， laying the foundation for the development of a diagnostic scale for qi stagnation syndrome.

Animals ; Mice ; DNA Repair ; DNA Breaks, Double-Stranded ; DNA/metabolism* ; DNA Damage

Objective To establish a preliminary evaluation system for gastrointestinal qi stagnation syndrome.Methods On the basis of the systematic evaluation of medical literature in the early stage of the research group,24 high-frequency items were subjected to Delphi method,the item indexes were determined through three rounds of expert consultation,and the proportion value of the indexes was determined by AHP,and the evaluation system of gastrointestinal qi stagnation syndrome was initially constructed.Results A total of 84 valid questionnaires were collected by three rounds of Delphi method,including 15 in the first round,32 in the second round and 37 in the third round.According to the statistics,16 items including distention(stuffy)or distending pain or moving pain(epigastric,abdominal,etc.),belching,borborygmus,flatus,etc.were selected.The order of the proportion of the first level indexes obtained by the analytic hierarchy process from high to low is:clinical symptoms,pulse,tongue;The proportion of secondary indicators from high to low is as follows:distention(stuffy)or distending pain or moving pain(epigastric,abdominal,etc.),pulse string,greasy fur,thin white fur,slippery pulse string,pulse sinking string,light red tongue,flatus,borborygmus,belching,induced or aggravated in case of emotional distress,hiccup,abdominal mass,anorexia,vomiting,belching and swallowing acid.Conclusion Delphi method and analytic hierarchy process have been used to study gastrointestinal qi stagnation syndrome,and an evaluation system has been preliminarily formed.The index structure is reasonable,targeted and has strong clinical practicability.

Objective To develop a nomogram based on pulmonary nodules preoperative CT signs and 3D quantitative parameters for predicting mediastinal lymph node metastases in patients with clinical stage ⅠA lung adenocarcinoma.Methods The imaging data of 164 patients who underwent preoperative CT scan and systematic lymph node dissection were analyzed retrospectively.Commercially available AI software was used to extract 3D quantitative parameters of pulmonary nodules automatically,and CT signs of pulmonary nodules were analyzed.Logistic regression was used to explore the role of these parameters in predicting pathological nodal involvement.A nomogram prediction model was established,then discrimination and calibration of the model were evaluated.Results Among 164 enrolled patients,19(11.6％)were tested positive for mediastinal lymph node metastases at pathology review.The nomogram incorporated spiculation,lobulation,the largest cross-sectional area,and carcinoembryonic antigen(CEA).The model showed great discrimination and calibration,with a C-index of 0.942[95％confidence interval(CI)0.923-0.961].The predicted value of the model fitted well with the actual observed value on the calibration curve.Conclusion The nomogram prediction model based on preoperative CT signs,3D quantitative parameters,and CEA can estimate the probability of mediastinal lymph node metastases in clinical stage ⅠA lung adenocarcinoma.This model may help with clinical decision-making and individualized evaluation.

The major reason for the resistance of Gram-negative bacteria to β-lactam antibiotics is the expression of β-lactamases.Metallo-β-lactamases (MBL) hydrolyze almost all types of β-lactam antibiotics including carbapenems, posing a challenge to global public health. Developing MBL inhibitors is an important method to treat the infections caused by resistant bacteria. As an important type of MBL inhibitors, chelating agents can inhibit MBL by chelating, stripping, and binding Zn2+ in the active center of MBL.This review summarizes recent publications on chelators as MBL inhibitors, discussing their chemical structures, inhibitory potency, synergistic effects with antibiotics, selectivity and mechanism of action, including EDTA and related compounds, aspergillomarasmine A (AMA) and its derivatives, NOTA and related compounds, pyridine carboxylic acid and pyridine methylamine compounds, aiming to provide reference for future development of potent, selective and safe clinical MBL inhibitors.

Chinese mitten crab (Eriocheir sinensis) is an important aquaculture species in Crustacea. Functional analysis, although essential, has been hindered due to the lack of sufficient genomic or transcriptomic resources. In this study, transcriptome sequencing was conducted on 59 samples rep-resenting diverse developmental stages (fertilized eggs, zoea, megalopa, three sub-stages of larvae, juvenile crabs, and adult crabs) and different tissues (eyestalk, hepatopancreas, and muscle from juvenile crabs, and eyestalk, hepatopancreas, muscle, heart, stomach, gill, thoracic ganglia, intes-tine, ovary, and testis from adult crabs) of E. sinensis. A comprehensive reference transcriptome was assembled, including 19,023 protein-coding genes. Hierarchical clustering based on 128 differ-entially expressed cuticle-related genes revealed two distinct expression patterns during the early lar-val developmental stages, demonstrating the distinct roles of these genes in 'crab-like"cuticle formation during metamorphosis and cuticle calcification after molting. Phylogenetic analysis of 1406 one-to-one orthologous gene families identified from seven arthropod species andCaenorhabditis elegans strongly supported the hypothesis that Malacostraca and Branchiopoda do not form a monophyletic group. Furthermore, Branchiopoda is more phylogenetically closely related to Hexapoda, and the clade of Hexapoda and Branchiopoda and the clade of Malacostraca belong to the Pancrustacea. This study offers a high-quality transcriptome resource for E. sinensis and demonstrates the evolutionary relationships of major arthropod groups. The differentially expressed genes identified in this study facilitate further investigation of the cuticle-related gene expression networks which are likely associated with'crab-like"cuticle formation during metamor-phosis and cuticle calcification after molting.

Tumor-specific neoantigens have attracted much attention since they can be used as biomarkers to predict therapeutic effects of immune checkpoint blockade therapy and as potential targets for cancer immunotherapy. In this study, we developed a comprehensive tumor-specific neoantigen database (TSNAdb v1.0), based on pan-cancer immunogenomic analyses of somatic mutation data and human leukocyte antigen (HLA) allele information for 16 tumor types with 7748 tumor samples from The Cancer Genome Atlas (TCGA) and The Cancer Immunome Atlas (TCIA). We predicted binding affinities between mutant/wild-type peptides and HLA class I molecules by NetMHCpan v2.8/v4.0, and presented detailed information of 3,707,562/1,146,961 potential neoantigens generated by somatic mutations of all tumor samples. Moreover, we employed recurrent mutations in combination with highly frequent HLA alleles to predict potential shared neoantigens across tumor patients, which would facilitate the discovery of putative targets for neoantigen-based cancer immunotherapy. TSNAdb is freely available at http://biopharm.zju.edu.cn/tsnadb.
Antigens, Neoplasm ; metabolism ; Data Analysis ; Databases, Genetic ; Humans ; Immunotherapy ; Mutation ; genetics ; Neoplasms ; genetics ; immunology ; Tumor Suppressor Protein p53 ; genetics ; Urinary Bladder Neoplasms ; genetics

Objective:To explore the proliferation inhibition and radiosensitization of Biyanqing granule on nasopharyngeal carci -noma cell line CNE-2 in vitro.Methods:CNE-2 cells were cultured in vitro.The inhibition of Biyanqing granule on the proliferation of CNE-2 cell was evaluated by MTT assay .Radiosensitization was explored by clone formation assay , and cell cycle and apopotosis were observed by flow cytometry ( FCM) .Results:Biyanqing granule could inhibit the proliferation of CNE-2 in a time-and dose-dependent manner.The IC50 in 24, 48 and 72 h was 70.79, 60.13 and 51.63 mg· ml-1(calculated according to the weight of all medicinal ma-terial), respectively.The colony formation assay showed that Biyanqing granule combined with radiation could significantly reduce the colony formation of CNE-2 cells.With the concentration increase of the main drug , the colony formation of CNE-2 cells was reduced . The number of colony formation in the negative control group , the radiation group , 10 mg· ml-1 and 20 mg· ml-1 Biyanqing combined with radiation groups (calculated according to the weight of all medicinal material ) was significant different (P<0.05).With the main drug concentration increasing , the percentage of G 2/M phase and apoptotic cells were both increased , and compared with the con-trol group, the difference was significant (P<0.05).Conclusion:Biyanqing granule can not only inhibit CNE-2 cells but also block CNE-2 cells in G2/M to improve the radiosensitization of CNE-2 cells.

Objective To explore the mechanism of six-segment classification of femur intertrochanteric fracture in a three-dimensional finite element model of upper thigh-bone and its clinical relevance.Methods The left upper thigh-bone of a normal male volunteer of 60 years old was scanned using 64-slice CT.After the images were stored in the format of JPG,they were input into photoshop 7.0 for three-dimensional creation.The three-dimensional images were used to make a three-dimensional finite element model of upper thigh-bone using software Super 93.The model consisted of 764 nodes and 531 units (including 306 compact bone units and 225 cancellous bone units).In this model,the stress distribution at the trochanter during human tumbling was analyzed by imitating adduction,abduction,adduction-intemal rotation and abduction-external rotation of the hip,as well as intense muscular contraction of gluteus medius,piriformis and iliopsoas during adduction-internal rotation of the hip.Results Analysis of stress nephogram showed that the stress was distributed mainly at the exterior cortical bone and spread to the inter-trochanteric part in simple hip adduction (two-part fracture) and distributed mainly at the interior cortical bone and spread to the inter-trochanteric part in simple hip abduction (two-part fracture).In adduction-internal rotation or abduction-external rotation of the hip,the stress was focused on the anterior and posterior walls of the greater trochanter and trochanteric part and spread to the exterior and interior cortical bone and the inter-trochanteric part (three-or four-part fracture).During intense muscular contraction of gluteus medius,piriformis and iliopsoas during adduction-internal rotation of the hip,the stress was focused on the anterior and posterior walls of the greater trochanter,lesser trochanter and trochanteric part and spread to the exterior and interior cortical bone and the inter-trochanteric part (five-or six-part fracture).Conclusions The six-segment classification of femur intertrochanteric fracture explains complex stress distributions after human falling.When combined with three-dimensional CT reconstruction,it is more intuitive than other classifications so that it can provide more definite surgical advice for different types of fracture.

Objective To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci ( Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods The fusion protein His-CPSIT_p7 was expressed in E.coli BL21 and purified by Ni-NTA affinity chromatography .BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CPSIT_p7 by ELISA.Penicillin sodium was used to establish a model of Cps persistence infection .RT-PCR and Western blot assay were performed to de-tect the expression of CPSIT_p7 at mRNA and protein levels during Cps persistent infection .Results The fusion protein His-CPSIT_p7 was successfully expressed with the use of constructed recombinant expression plasmid pET30 a-CPSIT_p7 and purified .ELISA result showed that the specific antibody titer against CPSIT_p7 reached 1 ∶1 000 000 on the 40th days after immunization .The expression of CPSIT_p7 at mRNA and protein levels were increased in a time-dependent manner in Cps-infected HeLa cells .The peak of mRNA level was reached at the time point of 36 hours after infection , followed by a time-dependent decrease during Cps acute infection .However , the expression of CPSIT_p7 at mRNA and protein levels were not decreased until 60 hours after infection during Cps persistent infection .Conclusion His-CPSIT_p7 protein was suc-cessfully expressed in the prokaryotic expression system and purified , showing an advantage of good immuno-genicity.Highly expressed CPSIT_p7 at mRNA and protein levels were detected during Cps persistent infection.