Aim To study the application status of odds ratio for medical animal experiments.Methods Odds ratio and seven kinds of animal models were used as retrieval strategy to search medical animal experiment related papers in several Chinese and English databases.Papers relating to each kind of animal model and using odds ratio in abstract and text were counted. Data from different databases were compared. Calculation of odds ratio was exemplified and the significance of different odds ratio values was illustrated in this paper.Results Few medical animal experiments cited odds ratio as statistics.Conclusions The importance of odds ratio has not been fully recognized in Chinese references.

Objective:To investigate the risk factors of central lymph node metastasis (CLNM) in cN0 paillary thyroicl microcarcinoma (PTMC) and to establish a nomogram model for predicting the probability of cN0 PTMC CLNM.Methods:The clinicopathological data of 192 patients with cN0 PTMC admitted to the Department of General Surgery of the Second Hospital of Lanzhou University from Aug. 2016 to Aug. 2020 were retrospectively analyzed. There were 41 males and 151 females, 50 with CLNM and 142 without CLNM. The patients were divided into 2 groups according to the presence or absence of pathologically confirmed CLNM. Patient’s age, gender, tumor diameter, multiple, with Hashimoto’s disease, with nodular goiter, with or without near the posterior dorsal membrane, aspect ratio >1, with or without extratumoral infiltration, with or without lymphadenopathy, TSH levels, and TG levels were statistically analyzed. Pearson chi-square test was used to analyze the count data of hypothesis test, and the R language software package was used for Logistic multivariate analysis. The entry conditions were screened by stepwise regression to establish a nomogram prediction model, and the Bootstrap method was used for model verification. P<0.05 was considered statistically significant. Results:Multivariate logistic analysis showed that extratumoral invasion ( P=0.032) , presence of lymphadenopathy ( P=0.010) , and TG>68 μg/L ( P=0.007) were risk factors for central lymph node metastasis. The optimal model was established by stepwise regression. The factors included tumor diameter ≥0.5 cm, nodular goiter, extratumoral invasion, lymphadenopathy and TG>68 μg/L (AIC: 212.27) . The nomogram model was established according to the above risk factors. The consistency index (c-index) was 0.711. The results of calibration graph drawing and internal and external validation demonstrated its good consistency and applicability. Conclusion:Extratumoral invasion, lymphadenopathy, and TG>68 μg/L are risk factors for cN0 PTMC CLNM, and the nomogram established in the study can effectively predict the CLNM rate in patients with cN0PTMC and contribute to clinicians’ diagnosis and treatment decisions.

Objective To clone and characterize Profilin encoding genes in Amaranthus spinosus and to analyze the contribution of different amino acids in isoallergens to allergen antigenicity and tertiary structure. Methods The primers were designed according to the core sequences which were obtained by bioinformatic analysis of the known Profilin amino acid sequences, followed by gene cloning from the Ama- ranthus spinosus cDNA pool and subsequent confirmation by double-digestion, colony PCR and DNA sequen- cing. Antigenicity evaluation and tertiary structural modeling of the encoded protein were accomplished by online software MULTIPRED and SWISS-MODEL, respectively. Results Two panallergenic genes, named as PRF7 and PRF23, were acquired from Amaranthus spinosus. Sequence and structure analysis demonstra- ted that there was some discrepancy in tertiary structures of the encoded proteins, besides distinct difference in their amino acid sequences. PRF7 exhibited high homology with panallergen Profilins Q64LH0, with the identities 98%, whereas the homology of PRF23 and Q9XF42 (apple allergen) was 81%. Q64LH0 and PRF23 were modeled as 3nulA (Q42449) and lg5uB (Q9LE18), respectively. PRF23 exhibited distinct0 three dimensional structural difference in certain fragments compared with Q64LH0 and other Profilins. Though the average values of antigenicity displayed no difference between Q64LH0 and PRF23 on whole se- quences, the antigenicity of PRF23 on certain fragments was obviously lower than that of Q64LHO because of the alteration of some amino acids with different characters, implying the cause of lower incidence of hay fe- ver in South China than in North China. Conclusion Based on sequence analysis, antigenicity evaluation and tertiary structural modeling for Q64LH0 and PRF23, we obtained lots of useful information about the contribution of different amino acids to antigenicity and protein structures, thus would facilitate allergen ge- netic improvement by amino acid replacement.

Objective To determine the relationship between early postoperative delirium (EPD) and prognosis in patients undergoing non-cardiac surgery. Methods This was a prospective cohort study consisted of 698 patients admitted to postanesthesia care unit, undergoing non-cardiac surgery under general anesthesia, between June and December 2009. The risk factors affecting prognosis were collected. All the patients were assessed for the development of delirium by experienced research staff using Confusion Assessment Method for Intensive Care Unit. The patients were divided into 2 groups according to the occurrence of EPD: EPD group and no EPD (NEPD) group. The postoperative hospital length of stay was made as a major prognostic indicator. Cox proportional hazard regression model was used to analyze the risk factors affecting prognosis. Results Of the 698 patients, 197 (28.2%) developed EPD. The postoperative hospital length of stay was prolonged in group EPD compared with group NEPD. The Cox proportional hazard regression model analysis indicated that EPD was an independent risk factor affecting prognosis. Conclusion EPD is an independent risk factor affecting prognosis in patients undergoing non-cardiac surgery.

Objective:To obtain purified recombinant Litopenaeus vannamei allergen protein Lit v 1.2.Methods: The target gene of Lit v 1.2 was inserted into clone vector pGEM-T and then ligated to the expression vector pET 44a.The pET44a-Liv 1.2 was transformed into Rosetta and screened by ampicillin resistance .The recombinant protein was expressed by IPTG induction .The protein was purified by 6-His tag affinity chromatography and the purification was analyzed by SDS-PAGE gel electrophoresis .Results:The ex-pression plasmid pET44a-Lit v 1.2 was constructed.SDS-PAGE showed that expressed Lit v 1.2 was efficient and soluble in E.coli Rosetta.The protein molecular weight was consistent with the theoretical value .The highly purified target protein was obtained.Conclusion:In this study ,we successfully gained highly purified recombinant allergen protein Lit v 1.2 which was expressed in prokaryotic system and purified by affinity chromatography column .The purified Lit v 1.2 protein will facilitate us to further study its role in immunological responses .

Objective:To develop and validate a clinical prediction model for the risk of malignant ventricular arrhythmia in patients with acute myocardial infarction (AMI) during hospitalization, and evaluate the effect of the prediction model.Methods:A retrospective study was conducted. A total of 2 649 patients with AMI admitted to cardiology department of Changzhou No.2 People's Hospital of Nanjing Medical University from December 2012 to August 2020 were enrolled. The clinical characteristics including gender, age, medical history, discharge diagnosis, vital signs during hospitalization, electrocardiogram characteristics at admission, laboratory examination indexes, interventional treatment, drug usage, malignant ventricular arrhythmias [mainly included sustained ventricular tachycardia (VT), ventricular flutter or ventricular fibrillation (VF)], and death were recorded. All patients were divided into two groups according to whether VT/VF occurred during their hospitalization. Independent risk factors for VT/VF during hospitalization were evaluated by multivariate Logistic regression analysis, and a clinical prediction model was constructed. The receiver operating characteristic curve (ROC curve) was plotted, and the area under ROC curve (AUC) was calculated to evaluate the accuracy of the prediction model.Results:A total of 2 649 eligible patients with AMI were enrolled, of whom 134 (5.06%) developed VT/VF during hospitalization. The in-hospital mortality rate in VT/VF group was significantly higher than that in non-VT/VF group (38.1% vs. 1.7%, P < 0.01). Compared with the non-VT/VF group, the patients in the VT/VF group with lower systolic blood pressure [SBP (mmHg, 1 mmHg = 0.133 kPa): 125.9±28.2 vs. 132.0±24.2], higher random blood glucose (mmol/L: 8.6±4.8 vs. 7.4±3.7), worse cardiac function [Killip heart function grade ≥ 3: 36.6% vs. 10.7%, left ventricular ejection fraction (LVEF) < 0.50: 56.7% vs. 33.6%, frequent premature ventricular contractions: 12.7% vs. 1.2%] and more hypokalemia (46.3% vs. 17.3%), with significant differences (all P < 0.05). Multivariate Logistic regression analysis showed that Killip classification of cardiac function ≥ 3 [odds ratio ( OR) = 3.540, 95% confidence interval (95% CI) was 2.336-5.363], random blood glucose > 11.1 mmol/L ( OR = 1.841, 95% CI was 1.171-2.893), LVEF < 0.50 ( OR = 0.546, 95% CI was 0.374-0.797), frequent premature ventricular contractions ( OR = 12.361, 95% CI was 6.077-25.144), potassium < 3.5 mmol/L ( OR = 4.268, 95% CI was 2.910-6.259), SBP < 90 mmHg ( OR = 0.299, 95% CI was 0.150-0.597) and creatinine (Cr) > 100 μmol/L ( OR = 2.498, 95% CI was 1.170-5.334) were independent risk factors for VT/VF in patients with AMI (all P < 0.05). The clinical prediction model of VT/VF risk was constructed based on the variables selected by multivariate regression analysis. The ROC curve analysis showed that the AUC of the model in predicting VT/VF was 0.779 (95% CI was 0.735-0.823, P < 0.001); the optimal cut-off value of the model was 17, the sensitivity was 76.1%, the specificity was 67.3%. Conclusions:The incidence of VT/VF during hospitalization of AMI patients significantly increases the risk of in-hospital death. The independent risk factors of VT/VF are Killip grade ≥ 3, random blood glucose > 11.1 mmol/L, LVEF < 0.50, frequent ventricular premature beats, potassium < 3.5 mmol/L, SBP < 90 mmHg and Cr > 100 μmol/L. The newly constructed clinical prediction model has certain predictive value for the occurrence risk of VT/VF.

Triple-negative breast cancer (TNBC) is one of the most aggressive subtypes of breast cancer, accounting for about 15%-20% of all breast cancer patients. Its characteristics such as young age of onset, high risk of recurrence and visceral metastasis, and poor prognosis make it an urgent problem to be solved in clinical practice. With the rapid progress of genomics and proteomics, more and more potential therapeutic targets of TNBC have been identified, and the preliminary results show that these targets are promising to transform the treatment mode of TNBC in the future. This article reviews the clinical trials and applications of TNBC targeted drugs, as well as gives a brief introduction to nanomedicine.

PURPOSE: Cell-in-cell structures are created by one living cell entering another homotypic or heterotypic living cell, which usually leads to the death of the internalized cell, specifically through caspase-dependent cell death (emperitosis) or lysosome-dependent cell death (entosis). Although entosis has attracted great attention, its occurrence is controversial, because one cell line used in its study (MCF-7) is deficient in caspase-3. METHODS: We investigated this issue using MCF-7 and A431 cell lines, which often display cell-in-cell invasion, and have different levels of caspase-3 expression. Cell-in-cell death morphology, microstructures, and signaling pathways were compared in the two cell lines. RESULTS: Our results confirmed that MCF-7 cells are caspase-3 deficient with a partial deletion in the CASP-3 gene. These cells underwent cell death that lacked typical apoptotic properties after staurosporine treatment, whereas caspase-3-sufficient A431 cells displayed typical apoptosis. The presence of caspase-3 was related neither to the lysosome-dependent nor to the caspase-dependent cell-in-cell death pathway. However, the existence of caspase-3 was associated with a switch from lysosome-dependent cell-in-cell death to the apoptotic cell-in-cell death pathway during entosis. Moreover, cellular hypoxia, mitochondrial swelling, release of cytochrome C, and autophagy were observed in internalized cells during entosis. CONCLUSION: The occurrence of caspase-independent entosis is not a cell-specific process. In addition, entosis actually represents a cellular self-repair system, functioning through autophagy, to degrade damaged mitochondria resulting from cellular hypoxia in cell-in-cell structures. However, sustained autophagy-associated signal activation, without reduction in cellular hypoxia, eventually leads to lysosome-dependent intracellular cell death.
Apoptosis ; Autophagy ; Caspase 3* ; Cell Death ; Cell Hypoxia ; Cell Line ; Cytochromes c ; Entosis ; MCF-7 Cells* ; Mitochondria ; Mitochondrial Swelling ; Staurosporine

Purpose: Accumulating evidence has suggested that toll-like receptor 4 (TLR4) is critically involved in the pathogenesis of asthma. The aim of this study was to investigate the role of TLR4 in toluene diisocyanate (TDI)-induced allergic airway inflammation. Methods: TLR4−/− and wild-type (WT) C57BL/10J mice were sensitized and challenged with TDI to generate a TDI-induced asthma model. B-cell lymphoma 2 (Bcl-2) inhibitors, ABT-199 (4 mg/kg) and ABT-737 (4 mg/kg), were intranasally given to TDI-exposed TLR4−/− mice after each challenge. Results: TDI exposure led to increased airway hyperresponsiveness (AHR), granulocyte flux, bronchial epithelial shedding and extensive submucosal collagen deposition, which were unexpectedly aggravated by TLR4 deficiency. Following TDI challenge, TLR4−/− mice exhibited down-regulated interleukin-17A and increased colony-stimulating factor 3 in bronchoalveolar lavage fluid (BALF), while WT mice did not. In addition, TLR4 deficiency robustly suppressed the expression of NOD-like receptor family pyrin domain containing 3 and NLR family CARD domain containing 4, decreased caspase-1 activity in TDI-exposed mice, but had no effect on the level of high mobility group box 1 in BALF. Flow cytometry revealed that TDI hampered both neutrophil and eosinophil apoptosis, of which neutrophil apoptosis was further inhibited in TDI-exposed TLR4−/− mice, with marked up-regulation of Bcl-2. Moreover, inhibition of Bcl-2 with either ABT-199 or ABT-737 significantly alleviated neutrophil recruitment by promoting apoptosis. Conclusions These data indicated that TLR4 deficiency promoted neutrophil infiltration by impairing its apoptosis via up-regulation of Bcl-2, thereby resulting in deteriorated AHR and airway inflammation, which suggests that TLR4 could be a negative regulator of TDI-induced neutrophilic inflammation.

Objective:To investigate the effect of preoperative hemoglobin (Hb) level on the risk of developing acute kidney injury (AKI) after percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI).Methods:A retrospective study was conducted. The hospitalized patients diagnosed with AMI who underwent PCI from May 2015 to May 2020 in the department of cardiology in the Affiliated Changzhou No.2 People's Hospital of Nanjing Medical University were enrolled. According to the serum creatinine (SCr) level before and after interventional therapy, the patients were divided into an AKI group and a non-AKI group. The difference in patients' Hb levels between the AKI and non-AKI groups was compared. Univariate and multivariate Logistic regression analyses were used to analyze the effects of Hb levels on the risk of AKI after interventional therapy in patients with AMI. Kaplan-Meier survival curve was used to evaluate the effects of Hb levels on patients with AMI in all-cause death in the hospital.Results:A total of 922 AMI patients were enrolled in this study, of which 165 patients (17.9%) developed AKI. Compared with the non-AKI group, female patients in the AKI group had a higher proportion [35.8% (59/165) vs. 26.9% (204/757)], older (age: 69.78±14.56 vs. 66.61±13.44), with a lower rate of smoking [42.4% (70/165) vs. 51.7% (391/757)] and a higher prevalence of hypertension [73.3% (121/165) vs. 63.5% (481/757)], however, the patients in AKI group also had a worse cardiac function [the proportion of Killip grade 3 or above was higher: 33.9% (56/165) vs. 13.9% (105/757)], lower Hb level (g/L: 127.61±22.18 vs. 132.79±19.45), and there were less patients using angiotensin converting enzyme inhibitor/angiotensin Ⅱreceptor blocker [ACEI/ARB, 60.0% (99/165) vs. 74.5% (564/757)] and more patients using diuretics [24.8% (41/165) vs. 17.7% (134/757)] in AKI group, the differences were statistically significant (all P < 0.05). Compared with non-AKI group, patients in AKI group had a longer operation time [operation time > 60 minutes: 4.2% (7/165) vs. 1.5% (11/757)] and received more contrast media during the operative procedure [contrast media > 100 mL: 16.4% (27/165) vs. 3.6% (27/757)], the individuals had a higher rate of intra-operative hypotension [16.4% (27/165) vs. 8.2% (62/757)], and more patients were implanted more than 2 stents [8.5% (14/165) vs. 3.6% (27/757), all P < 0.05]. Univariate Logistic regression analysis suggested that each 1 g/L increase in preoperative Hb level was associated with a 1.2% decrease in the risk of postoperative AKI [odds ratio ( OR) = 0.988, 95% confidence interval (95% CI) was 0.980-0.996, P = 0.003]. Meanwhile, for every 1 standard deviation increase in preoperative Hb level, the risk of postoperative AKI decreased by 22.1% ( OR = 0.779, 95% CI was 0.661-0.918, P = 0.003). The patients were divided into low, medium and high concentration groups according to Hb levels (Hb levels were < 110 g/L, 110-150 g/L, ≥ 150 g/L, respectively), and multivariate Logistic regression analysis showed that the risk of AKI was significantly reduced in the high concentration group compared with that in the low concentration group ( OR = 0.463, 95% CI was 0.241-0.888, P = 0.020). The Kaplan-Meier survival curve analysis indicated that the short term survival after coronary intervention in AMI patients with low Hb concentration was significantly lower than that in patients with medium and high Hb concentration (Log-Rank: χ2= 23.215, P < 0.001). Conclusions:Preoperative lower Hb level is an independent risk factor for postoperative AKI in AMI patients. AMI patients with lower Hb levels have an increased risk of all-cause mortality within 1 month after AMI.