[Nomogram-Based Prediction of Risk of Cervical Lymph Node Metastasis in Differentiated Thyroid Carcinoma].

Objective To establish a nomogram for predicting the risk of cervical lymph node metastasis in differentiated thyroid carcinoma (DTC). Methods The patients with complete clinical data of DTC and cervical lymph node ultrasound and diagnosed based on pathological evidence from January 2019 to December 2021 were assigned into a training group (n=444) and a validation group (n=125).Lasso regression was performed to screen the data with differences between groups,and multivariate Logistic regression to establish a prediction model with the factors screened out by Lasso regression.C-index and calibration chart were employed to evaluate the prediction performance of the established model. Results The predictive factors for establishing the model were lymph node short diameter≥0.5 cm,long-to-short-axis ratio<2,disappearance of lymph node hilum,cystic transformation,hyperechogenicity,calcification,and abnormal blood flow (all P<0.001).The established model demonstrated a good discriminative ability,with the C index of 0.938 (95%CI=0.926-0.961) in the training group. Conclusion The nomogram established based on the ultrasound image features of cervical lymph nodes in DTC can accurately predict the risk of cervical lymph node metastasis in DTC.

[Influencing Factors and Prediction Model of Performance of Needle Visualization in Fine Needle Aspiration of Thyroid Nodules].

Objective To investigate the influencing factors and establish a model predicting the performance of needle visualization in fine-needle aspiration (FNA) of thyroid nodules. Methods This study prospectively included 175 patients who underwent FNA of thyroid nodules in the Department of Ultrasound in China-Japan Friendship Hospital and compared the display of the needle tips in the examination of 199 thyroid nodules before and after the application of needle visualization.We recorded the location,the positional relationship with thyroid capsule,ultrasonic characteristics,and the distribution of the soft tissue strip structure at the puncture site of the nodules with unclear needle tips display before using needle visualization.Furthermore,according to the thyroid imaging reporting and data system proposed by the American College of Radiology,we graded the risk of the nodules.Lasso-Logistic regression was employed to screen out the factors influencing the performance of needle visualization and establish a nomogram for prediction. Results The needle tips were not clearly displayed in the examination of 135 (67.8%) and 53 (26.6%) nodules before and after the application of needle visualization,respectively,which showed a significant difference (P<0.001).Based on the positional relationship between the nodule and capsule,anteroposterior/transverse diameter (A/T) ratio,blood supply,and the distribution of subcutaneous strip structure at the puncture site,a nomogram was established to predict the probability of unclear display of the needle tips after application of needle visualization.The C-index of the prediction model was 0.75 (95%CI=0.67-0.84) and the area under the receiver operating characteristic curve was 0.72.The calibration curve confirmed the appreciable reliability of the prediction model,with the C-index of 0.70 in internal validation. Conclusions Needle visualization can improve the display of the needle tip in ultrasound-guided FNA of thyroid nodules.The nomogram established based on ultrasound features such as the positional relationship between the nodule and capsule,A/T ratio,blood supply,and the distribution of subcutaneous strip structure at the puncture site can predict whether needle visualization is suitable for the examination of nodules.

Quantitative detection of parasitic ciliate Cryptocaryon irritans in seawater with an optimized sample processing method.

The protozoan Cryptocaryon irritans is one of the most important ectoparasites of marine fish, causing 'white spot disease' and mass mortality in aquaculture. To accurately predict disease outbreaks and develop prevention strategies, improved detection methods are required that are sensitive, convenient and rapid. In this study, a pair of specific primers based on the C. irritans 18S rRNA gene was developed and used in a real-time PCR (qPCR) assay. This assay was able to detect five theronts in 1 L of natural seawater. Furthermore, a linear model was established to analyse the log of Ct value and parasite abundance in seawater (y = -2.9623x + 24.2930), and the coefficient of determination (R2 ) value was 0.979. A lysis buffer was optimized for theront DNA extraction and used for storage sample. This method was superior to the commercial water DNA kit, and there was no significant degradation of DNA at room temperature for 24-96 hr. A dilution method was developed to manage qPCR inhibitors and used to investigate natural seawater samples in a net cage farm with diseased fish, and the findings were consistent with the actual situation. This study provides a valuable tool for assisting in the early monitoring and control of cryptocaryoniasis in aquaculture.

[Application of Ultrasonography in the Diagnosis of Medullary Thyroid Cancer].

Despite its low incidence,medullary thyroid carcinoma(MTC)is featured by its fast progression and poor prognosis.Early diagnosis and treatment is therefore particularly important.As a convenient and non-invasive diagnostic tool,ultrasound plays a key role in the diagnosis and follow-up of MTC.In recent years,the application of conventional ultrasound,ultrasonic elastography,and ultrasound-guided fine needle aspiration biopsy has dramatically improved the diagnostic accuracy of MTC.

[Start-up and Process Characteristics of Simultaneous ANAMMOX and Denitrification (SAD) in a Pilot-scale Anaerobic Sequencing Batch Reactor (ASBR)].

A pilot scale anaerobic sequencing batch reactor (ASBR, working volume 530 L), inoculated by anaerobic sludge from an A2O process, was developed to investigate the start-up of anaerobic ammonia oxidation (ANAMMOX) and its combination with denitrification for deep-level nitrogen removal from saline wastewater. Simultaneously, the flora structure was analyzed. Results showed that under the conditions of temperature 35℃±1℃ and reaction time 14 h, ANAMMOX was successfully started-up after 160 days of operation. During the stabilized operation stage, ANAMMOX coupled with denitrification (SAD) led to a total nitrogen (TN) removal efficiency and removal rate of 91.1% and 0.45 kg·(m3·d)-1, respectively. The successful cultivated sludge formed granules and presented as a light red color, with the main bacteria genus being Candidatus Brocadia (10.6%). Additionally, high efficiency nitrogen and organic carbon removal (COD and TN removal efficiency of 93.2% and 90.0%, respectively) from wastewater simulating desulfurization and denitrification tailings with high salinity (Cl- concentration of 8000 mg·L-1) was achieved in the SAD system by gradually increasing the salinity gradient. Moreover, the denitrification in SAD was mostly NO3--N→N2, with partial denitrification (NO3--N→NO2--N) accounting for only 30.3%.

[Effect of Salinity on Nitrogen Removal Performance of a Pilot-scale Anaerobic Ammonia Oxidation Process and Its Recovery Kinetics].

The denitrification characteristics of anaerobic ammonia oxidizing bacteria (AnAOB) treating high salinity wastewater were investigated in an pilot-scale anaerobic sequencing batch reactor (ASBR, 530 L) by gradually increasing the Cl- concentration. The results showed that AnAOB can adapt to the high salinity (Cl- concentration of 10000 mg·L-1) environment for high-efficiency denitrification by means of gradual salinity acclimation and total nitrogen (TN) removal rate of up to 92.3%. In particular, the denitrification performance was influenced by two gradients of Cl- concentrations, namely 6000 mg·L-1 and 10000 mg·L-1, but it could be gradually recovered as the acclimatization process continued. The modified Boltzmann model accurately fit the activity recovery process of AnAOB after being inhibited by the different salinities, and the correlation coefficient R2 was above 0.96. The fitted recovered median values tc for Cl- concentrations of 6000 mg·L-1 and 10000 mg·L-1 were 28.765 d and 44.495 d. NRRmax for these concentrations was 0.145 kg·(m3·d)-1 and 0.212 kg·(m3·d)-1, and NRRmin was 0.021 kg·(m3·d)-1 and 0.085 kg·(m3·d)-1, respectively. After salinity acclimation, the dominant bacteria of AnAOB were Candidatus Brocadia and Candidatus Jettenia (the abundances were 14.76% and 2.7%, respectively), the granulation degree and sludge density increased to varying degrees, and the sludge color was reddish brown.

[Suppression and Recovery Characteristics of Pilot-scale ANAMMOX-ASBR System Treating Desulfurization and Denitrification Tailings from Thermal Power Plant].

A pilot-scale anaerobic sequencing batch reactor (ASBR, working volume 530 L), inoculated with oxygen-segmented sludge in an oxidation ditch process, was developed to investigate the start-up of anaerobic ammonium oxidation (ANAMMOX) and its combination with denitrification for deep-level nitrogen removal from desulfurization and denitrification tailings of a thermal power plant. The results showed that, under conditions with a temperature of (35±1)℃and reaction time of 20 h, ANAMMOX was successfully started up after 180 days. During the stable operations phase, total nitrogen (TN) removal rate and removal efficiency reached 91.1% and 0.3 kg·(m3·d)-1, respectively. During the activity suppression stage of the ANAMMOX-ASBR treating real desulfurization and denitrification tailings, the recovery of its activity could be achieved in 93 days by removing inhibitory factors (Cl- concentration) and reducing the concentration of influent substrate. In addition, by gradually increasing the addition ratio of desulfurization and denitrification tails (30%, 70%, and 100%), the coupling of ANAMMOX and denitrification was achieved in the ASBR to ensure stable effluent TN removal rate and COD concentrations below 92% and 88.5 mg·L-1, respectively. The modified logistic model was more suitable for the NRR recovery process after ANAMMOX was impacted by desulfurization and denitrification tailings. The NRR recovery delay time λ was 17.777 cycles, the and R2 was 0.92948.

Abdominal paracentesis drainage ameliorates severe acute pancreatitis in rats by regulating the polarization of peritoneal macrophages.

AIM: To investigate the role of peritoneal macrophage (PM) polarization in the therapeutic effect of abdominal paracentesis drainage (APD) on severe acute pancreatitis (SAP). METHODS: SAP was induced by 5% Na-taurocholate retrograde injection in Sprague-Dawley rats. APD was performed by inserting a drainage tube with a vacuum ball into the lower right abdomen of the rats immediately after the induction of SAP. To verify the effect of APD on macrophages, PMs were isolated and cultured in an environment, with the peritoneal inflammatory environment simulated by the addition of peritoneal lavage in complete RPMI 1640 medium. Hematoxylin and eosin staining was performed. The levels of pancreatitis biomarkers amylase and lipase as well as the levels of inflammatory mediators in the blood and peritoneal lavage were determined. The polarization phenotypes of the PMs were identified by detecting the marker expression of M1/M2 macrophages via flow cytometry, qPCR and immunohistochemical staining. The protein expression in macrophages that had infiltrated the pancreas was determined by Western blot. RESULTS: APD treatment significantly reduced the histopathological scores and levels of amylase, lipase, tumor necrosis factor-α and interleukin (IL)-1ß, indicating that APD ameliorates the severity of SAP. Importantly, we found that APD treatment polarized PMs towards the M2 phenotype, as evidenced by the reduced number of M1 macrophages and the reduced levels of pro-inflammatory mediators, such as IL-1ß and L-selectin, as well as the increased number of M2 macrophages and increased levels of anti-inflammatory mediators, such as IL-4 and IL-10. Furthermore, in an in vitro study wherein peritoneal lavage from the APD group was added to the cultured PMs to simulate the peritoneal inflammatory environment, PMs also exhibited a dominant M2 phenotype, resulting in a significantly lower level of inflammation. Finally, APD treatment increased the proportion of M2 macrophages and upregulated the expression of the anti-inflammatory protein Arg-1 in the pancreas of SAP model rats. CONCLUSION: These findings suggest that APD treatment exerts anti-inflammatory effects by regulating the M2 polarization of PMs, providing novel insights into the mechanism underlying its therapeutic effect.

Curcumin cytotoxicity is enhanced by PTEN disruption in colorectal cancer cells.

AIM: To investigate the effects of phosphatase and tensin homolog deleted on chromosome 10 (PTEN) deficiency on the cytotoxicity of chemotherapeutic agents toward colorectal cancer cells. METHODS: PTEN-deficient colorectal cancer (CRC) cells were generated by human somatic cell gene targeting using the adeno-associated virus system. The cytotoxic effects of compounds including curcumin, 5-fluorouracil (5-FU), dihydroartemisinin (DHA), irinotecan (CPT-11) and oxaliplatin (OXA) on cancer cells were determined using the MTT assay. Enhanced cytotoxicity of curcumin in PTEN-deficient CRC cells was observed, and this was confirmed using clonogenic assays. Apoptosis and cell cycle progression were analyzed by flow cytometry. Levels of apoptosis and cell cycle-related proteins were examined by Western blotting. RESULTS: We developed an isogenic set of CRC cell lines that differed only in their PTEN status. Using this set of cell lines, we found that disruption of the PTEN gene had no effect on the sensitivity of CRC cells to 5-FU, CPT-11, DHA, or OXA, whereas PTEN disruption increased the sensitivity of CRC cells to curcumin. Loss of PTEN did not alter the curcumin-induced apoptosis in CRC cells. However, PTEN deficiency led to an altered pattern of curcumin-mediated cell cycle arrest. In HCT116 PTEN (+/+) cells, curcumin caused a G2/M phase arrest, whereas it caused a G0/G1 phase arrest in HCT116 PTEN (-/-) cells. Levels of cell cycle-related proteins were consistent with these respective patterns of cell cycle arrest. CONCLUSION: Curcumin shows enhanced cytotoxicity toward PTEN-deficient cancer cells, suggesting that it might be a potential chemotherapeutic agent for cancers harboring PTEN mutations.

A colloidal gold probe-based silver enhancement immunochromatographic assay for the rapid detection of abrin-a.

High-affinity anti-abrin-a monoclonal and polyclonal antibodies were used to develop a sandwich immunochromatographic assay and silver enhancement technology was used to further increase the sensitivity. Using a matrix of double distilled water or soybean milk with added abrin-a, the visual detection limit was found to be 10 ng mL(-1). The detection limit was 0.1 ng mL(-1) for abrin-a, an increase in sensitivity of 100-fold when the silver enhancement technology was employed. The assay was portable and very simple to perform and the detection was completed within 20 min without complicated handling procedures. There was no significant cross-reactivity with several homologous toxins and associated agglutinin. The assay reagents could be stored for 12 weeks at 4°C without significant loss of activity. These characteristics make the strip assay to be an ideal candidate for the development of a rapid toxin detection kit.

Preparation and identification of monoclonal antibody against abrin-a.

BALB/c mice were immunized four times with formalin-prepared abrin-a. Using the polyethylene glycol method, immunized splenocytes were isolated and fused with SP2/0 cells. An indirect ELISA was established and used to detect positive clones secreting monoclonal antibodies (mAbs) against abrin-a. After analysis, three hybridoma clones secreting IgG-subtype mAbs were obtained. The antibodies were purified from the hybridoma growth medium using protein A or G affinity chromatography. Western blot analysis was used to analyze the antigenic epitopes on abrin-a recognized by the mAbs. The mAbs were specific for abrin-a, with no detectable cross-reactivity with several homologous toxins and associated agglutinins. Sandwich ELISA was then developed using these mAbs, which had a detection limit for abrin-a of 7.8 ng/mL.