Objective To obtain monoclonal antibodies ( mAbs ) against neutrophil gelatinase-associated lipocalin ( NGAL ) and a chemiluminescense immune quantification assay based one paired mAbs.Methods Six-to-eight weeks old female BALB/c mice were immunized with the purified recombinant human NGAL antigen( rhNGAL) that was produced by the Escherichia coil expression system.The spleen was fused with hybridoma for screening anti-NGAL monoclonal antibodies by indirect ELISA.Western blot was implemented to identify the reactivity with native NGAL. Results The rhNGAL antigen was found to form disulfide cross-linked dimers and present excellent immunogenicity.The reaction titer of the immune serum of NGAL immunized mice was about 106.Thirty mAbs were screened by indirect ELISA, hereinto;the EC50 values of mAb23C12 and 38D10 were 0.034 g/mL, 0.022 g/mL respectively.The antibodies pair, 38D10/23C12-SAE labeled with AcridiniumEster(AE), were shown to work well in chemiluminescense immune response quantitative detection which was screened by NGAL standardand clinical urine samples.This detection can resolve positive and negative samples with a statistically significant difference (P<0.0001).And the correlation coefficient R2between NGAL quantitative results and that of the Abbott's NGAL chemiluminescence immune assay kit was greater than 0.97.The detection linear range was 10-1500 ng/mL, analytical sensitivity of the method was 0.63 ng/mL.Conclusion Highly purified rhNGAL antigen and specific anti-NGAL monoclonal antibodies are generated in this study.The detection capability of method is comparable with that of the international commercial kit.

Objective To construct an intelligent analysis system based on visual attention for diabetic retinopathy ( DR) assistant diagnosis and to realize the automatic screening and grading of fundus images of DR. Methods Total of 35126 DR fundus images were downloaded from the Diabetic Retinopathy Detection competition in the Data Modeling and Data Analysis Competition Platform (Kaggle),and 1200 fundus images were downloaded from the Messidor website. Firstly,according to the characteristics of DR fundus images,a series of preprocessing was carried out for retina images. Then,on the basis of VGG16 network,visual attention SENet module was introduced to improve the saliency of lesion features,and a deep convolution neural network SEVGG with complex network structure was generated. The network basically inherited some structural parameters of VGG16,and the parameters of SENet module were adjusted according to the basic network and training data set. Finally, the SEVGG network model was used to screen the DR fundus image,and the fundus image was divided into different levels according to the degree of lesions of DR in different periods. Configure the training platform and environment and perform algorithm performance verification experiments. Results The method proposed in this study was tested on different open standard datasets,and finally achieved high accuracy in image-based classification. The accuracy of 5 classification in Kaggle dataset was 83％,the sensitivity of lesion detection was 99. 86％ and the specificity was 99. 63％. The accuracy rate of the 4 classification in the Messidor data set was up to 88％,the sensitivity of the lesion detection was 98. 17％,and the specificity was 96. 39％. The introduction of visual attention was more significant for the focus of the lesion,which helped the accurate detection of DR. Conclusions This method effectively avoids some shortcomings of traditional artificial feature extraction and fundus image classification,and is more accurate for lesion recognition. It is not only superior to the previous method,but also shows better robustness and generalization.

[Objective]To observe the clinical effect of"lung-intestinal treatment"method for adjuvant treatment of acute exacerbation of chronic obstructive pulmonary disease(AECOPD)mechanical ventilation patients with syndrome of lung heat and Fu-viscera excess.[Methods]Sixty-two patients with AECOPD in Pulmonary and Critical Care Medicine(PCCM)and Intensive Care Unit(ICU)of the Third Affiliated Hospital of Zhejiang Chinese Medical University were selected and divided into conventional treatment group and treatment group by random number table,31 cases in each group.The conventional treatment group was treated with western medicine,and the treatment group was added with lung-intestinal treatment on the basis of the conventional treatment group.The traditional Chinese medicine(TCM)syndrome points,blood gas index,ventilator parameters,mechanical ventilation time,complications and clinical effects were observed.[Results]Compared with before treatment,the conventional treatment group had improvement in the blood gas index,ventilator parameters,TCM syndrome(cough and asthma,sputum,abdominal distension and constipation,fever,chest tightness)and total points after treatment(P＜0.05);after treatment,the blood gas index,ventilator parameters,TCM syndrome(cough and asthma,sputum,abdominal distension and constipation,fever,chest tightness,loss of appetite)were relieved and total scores were all improved in the treatment group(P＜0.05).Compared with the conventional treatment group,the treatment group had significantly improved blood gas index,ventilator parameters and clinical effects,the TCM symptoms(chest tightness,cough and asthma,abdominal distension and constipation,loss of appetite)were relieved and total scores were improved(P＜0.05),and the time of mechanical ventilation and complications were reduced compared with the conventional treatment group(P＜0.05).[Conclusion]The treatment of AECOPD mechanical ventilation patients with"lung-intestinal treatment"method has clear cu rative effect,which can relieve clinical symptoms,shorten the time of mechanical ventilation,and the efficacy is better than that of western medicine alone.

Objective:To investigate the factors related to recurrent laryngeal nerve invasion in papillary thyroid carcinoma (PTC) with posterior capsular involvment.Methods:The data of 186 PTC patients admitted and operated from Jun 2017 to Jun 2022 were retrospectively analyzed. The invasion of recurrent laryngeal nerve was evaluated on its relation to gender, age, tumor size, Hashimoto's thyroiditis, lymph node metastasis in central region, BRAFV600E gene mutation especially PTC posterior capsular involvement.Results:The recurrent laryngeal nerve was invaded in 30 out of 186 patients. Univariate analysis showed that recurrent laryngeal nerve invasion was related to tumor size, Hashimoto's thyroiditis and cervical lymph node metastasis( χ2=6.964,4.814,6.078, P<0.05). Multivariate regression analysis showed that tumor size and lymph node metastasis in cervical region were independent risk factors for recurrent laryngeal nerve invasion(β=1.020,1.622, P<0.05). Hashimoto's thyroiditis was a protective factor for recurrent laryngeal nerve invasion (β=-1.881, P<0.05). Conclusions:When papillary thyroid carcinoma invaded the capsule, the risk of recurrent laryngeal nerve invasion was higher with larger tumor size and cervical lymph node metastasis, while Hashimoto's thyroiditis was a protective factor for the risk of recurrent nerve invasion.

Objective:To investigate the factors affecting the change of serum calcium and phosphorus after total parathyroidectomy for secondary hyperparathyroidism(SHPT).Method:Sixty patients with secondary hyperparathyroidism undergoing total parathyroidectomy were selected.Gender, age, hemodialysis time, bone pain time, intravenous calcium supplementation time, blood parathyroid hormone(PTH), parathyroid mass, blood calcium, phosphorus and alkaline phosphatase at admission were tested to compare the correlation between postoperative calcium and phosphorus changes.Result:Blood calcium was: (2.50±0.02), (2.27±0.03), (1.97±0.03), (1.95±0.03), (1.95±0.03), (2.03±0.02) mmol/L at admission, 6 hours after surgery, 1, 2, 3 days after surgery, and when discharged. Blood phosphorus was: (2.32±0.07), (1.95±0.05), (1.65±0.06), (1.44±0.05), (1.35±0.36), (1.14±0.04) mmol/L and when discharged.There are differences between pairwise comparisons ( F=100.979, P<0.05; F=118.910, P<0.05). The blood PTH was (1 791±760) pg/ml, and the postoperative intravenous calcium supplementation time was (7.13±2.34) days. Blood PTH was positively correlated with bone pain time, blood phosphorus and parathyroid mass at admission ( t=-2.199, P<0.05; t=4.718, P<0.05; t=2.507, P<0.05). The time of intravenous calcium supplementation was positively correlated with ALP at admission ( t=3.098, P<0.05). Preoperative serum PTH > 1 430 pg/ml or ALP > 191 mmol/L were prone to hypocalcemia on the first day after surgery. Conclusions:The longer bone pain time, the higher blood phosphorus and the more obvious parathyroid hyperplasia were, the higher PTH was. The higher the ALP, the longer the time of intravenous calcium supplementation. Preoperative serum PTH and ALP values can be used as predictors of hypocalcemia on the first day after surgery.

Objective To predict the causal relationship between immune cells and osteomyelitis(OM)by using Mendelian randomization.Methods Data sets of 731 immune cells and osteomyelitis were obtained from the UK Biobank and the genome wide association study.The inverse variance weighted method,weighted median method,and MR-Egger method were employed sequentially to verify the causal relationship between the 731 immune cells and osteomyelitis.Cochran's Q test,MR-Egger intercept test,mendelian randomization pleiotropy residual sum and outlier(MR-PRESSO),and leave-one-out analysis were evaluatede the stability and reliability of the results.Results A total of 21 immune cells were found to have a causal relationship with osteomyelitis(P＜0.05).Among them,T regulatory cells,classical dendritic cells and B cells were exhibited more stable causal relationships with osteomyelitis(P＜0.01).Specifically,T regulatory cells(OR=0.925,β=0.078)were identified as protective factors reducing the risk of osteomyelitis,while classical dendritic cells(OR=1.084,β=0.081)and B cells(OR=1.063,β=0.061)were identified as risk factors increasing the risk of osteomyelitis.Conclusion Twenty-one immune cells are genetically associated with the risk of osteomyelitis.

Objective:To investigate the incidence of venous thromboembolism (VTE) in patients with esophageal cancer (EC).Methods:The retrospective cohort study was conducted. The clinicopathological data of 8 458 EC patients who were admitted to Sichuan Cancer Hospital from January 2017 to December 2021 were collected. There were 6 923 males and 1 535 females, aged (64±9)years. There were 3 187 patients undergoing surgical treatment, and 5 271 cases undergoing non-surgical treatment. Observation indicators: (1) incidence of VTE in EC patients; (2) treatment and outcomes of patients with VTE. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was analyzed using the nonparameter rank sum test. Count data were expressed as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Comparison of ordinal data was analyzed using the nonparameter rank sum test. Results:(1) Incidence of VTE in EC patients. Of 8 458 EC patients, 175 cases developed VTE, with an incidence rate of 2.069%(175/8 458). Among 175 VTE patients, there were 164 cases of deep venous thrombosis (DVT), 4 cases of pulmonary embolism (PE), 7 cases of DVT and PE. There were 59 surgical patients and 116 non-surgical patients. There was no significant difference in thrombus type between surgical and non-surgical EC patients with VTE ( χ2=1.95, P>0.05). Of 3 187 surgical patients, the incidence of VTE was 1.851%(59/3 187), including an incidence of 0.157%(5/3 187) of PE. PE accounted for 8.475%(5/59) of surgical patients with VTE. Of 5 271 non-surgical patients, the incidence of VTE was 2.201%(116/5 271), including an incidence of 0.114%(6/5 271) of PE. PE accounted for 5.172%(6/116) of non-surgical patients with VTE. There was no significant difference in the incidence of VTE or PE between surgical patients and non-surgical patients ( χ2=1.20, 0.05, P>0.05). (2) Treatment and outcomes of patients with VTE. Among 175 EC patients with VTE, 163 cases underwent drug treatment, and 12 cases did not receive treatment. Among 163 cases with drug therapy, 158 cases underwent anticoagulant therapy, 5 cases were treated with thrombolysis. All the 163 patients were improved and discharged from hospital. Conclusions:The incidence of VTE in patients with EC is relatively low, as 2.069%. There is no significant difference in the incidence of VTE or thrombus type between surgical EC patients and non-surgical EC patients.