[Automatic classification of immune-mediated glomerular diseases based on multi-modal multi-instance learning].

OBJECTIVE: To develop a multi-modal deep learning method for automatic classification of immune-mediated glomerular diseases based on images of optical microscopy (OM), immunofluorescence microscopy (IM), and transmission electron microscopy (TEM). METHODS: We retrospectively collected the pathological images from 273 patients and constructed a multi-modal multi- instance model for classification of 3 immune-mediated glomerular diseases, namely immunoglobulin A nephropathy (IgAN), membranous nephropathy (MN), and lupus nephritis (LN). This model adopts an instance-level multi-instance learning (I-MIL) method to select the TEM images for multi-modal feature fusion with the OM images and IM images of the same patient. By comparing this model with unimodal and bimodal models, we explored different combinations of the 3 modalities and the optimal methods for modal feature fusion. RESULTS: The multi-modal multi-instance model combining OM, IM, and TEM images had a disease classification accuracy of (88.34±2.12)%, superior to that of the optimal unimodal model [(87.08±4.25)%] and that of the optimal bimodal model [(87.92±3.06)%]. CONCLUSION: This multi- modal multi- instance model based on OM, IM, and TEM images can achieve automatic classification of immune-mediated glomerular diseases with a good classification accuracy.

[Serum levels of endothelin-1 and connective tissue growth factor are elevated in patients with atrial fibrillation and correlated with relapse following radiofrequency ablation].

OBJECTIVE: To investigate the changes in serum levels of endothelin-1 (ET-1) and connective tissue growth factor (CTGF) in patients with atrial fibrillation (AF) and their value for predicting recurrence of AF after radiofrequency ablation (RFCA). METHODS: Sixty-six patients with paroxysmal AF (PaAF) and 72 with persistent AF (PaAF) admitted in our hospital were recruited as AF group and 80 patients with sinus rhythm as the control group, and in all the participants, serum levels of ET-1 and CTGF were measured using ELISA and Western blotting. From 6 patients with AF and 6 with sinus rhythm undergoing cardiac surgery in our hospital, tissue samples of the right atrial appendage were taken intraoperatively for observation of structural changes of the cardiomyocytes, myocardial fibrosis and expression of ET-1 and CTGF protein. In AF group, the patients receiving RFCA were followed up for 6 months following the procedure for assessment of the outcomes. RESULTS: Compared with the control patients, the patients with AF showed obvious damages of the cardiomyocyte structure and myocardial fibrosis. Serum levels of ET-1 and CTGF levels were significantly higher in PaAF and PeAF groups than in the control group, and were higher in PeAF group than in PaAF group. In the patients with AF, serum ET-1 and CTGF levels were positively correlated with left atrial diameter (LAD) (P < 0.05), and ET-1 was positively correlated with CTGF levels (P < 0.05). In patients with postoperative AF recurrence, the serum levels of ET-1 and CTGF were significantly higher than those in patients without recurrence; serum ET-1 and CTGF levels before and after the operation were positively correlated with the recurrence of PeAF, and elevated serum levels of ET- 1 and CTGF were identified by logistic regression analysis as independent risk factors for postoperative recurrence of PeAF. CONCLUSION: Serum levels of ET-1 and CTGF are significantly elevated in AF patients in positive correlation with AF duration. ET-1 and CTGF levels are higher in AF patients with postoperative recurrence, and they both have predictive value for recurrence of PeAF following RFCA.

[Correlation between the primary tumor size of endometrial carcinoma and lymph node metastasis and recurrence].

Objective: To investigate the clinical significance of the primary tumor size in patients with endometrial carcinoma (EC). Methods: A total of 385 patients with EC admitted to Peking University People's Hospital from January 2006 to December 2016 with complete follow up data were selected, whose tumor size data before biopsy were retrospectively studied. Results: (1) The mean diameter of the primary tumor was (3.6±1.8) cm (range: 1-15 cm). And 48 cases were 0-<2 cm, 78 cases were 2-<3 cm, 92 cases were 3-<4 cm, 73 cases were 4-<5 cm, 94 cases were ≥5 cm. The diameter of the tumor was associated with age <60 years old, premenopause, CA125≥35 kU/L, non-parturition, poor differentiation, stage Ⅲ-Ⅳ, depth of myometrial infiltration ≥1/2, cervical interstitial involvement, adnexal metastasis and lymph node metastasis (all P<0.05), but not associated with body mass index, hypertension, diabetes mellitus, pathology, lymph-vascular space invasion (all P>0.05). (2) Among the 334 patients underwent lymphadenectomy, 45 (13.5%, 45/334) cases with lymph node metastasis were observed. Stratified analysis showed that lymph node metastasis and recurrence rate of patients with EC gradually increased with the increase of tumor size (P<0.05). Adopting 2, 3, 4 and 5 cm as cut-off values of tumor size, there were significant differences in the rate of lymph node metastasis and recurrence among them observed (P<0.05), except for lymph node metastasis rate and recurrence rate when the cut-off value was 2 cm (P>0.05). (3) An receiver operating characteristic (ROC) curve analysis showed that a tumor diameter of 4.25 cm was the cut-off prognostic value to predict lymph node metastasis and recurrence of EC. Conclusions: Tumor diameter is significantly correlated with lymph node metastasis and recurrence in patients with EC. Tumor size should be considered in determining the scope of surgery and adjuvant therapy.

Expression and Clinical Significance of OX40 and OX40L mRNA in Hepatocellular Carcinoma.

In a retrospective study, the expression of mRNA of membrane receptor OX40 and its ligand OX40L in liver tissues was analyzed in 34 patients with hepatocellular carcinoma in order to assess their clinical implications and prognostic value. Expression of mRNA was analyzed by reverse transcription PCR and TaqMan probes. Expression of OX40 mRNA was significantly higher in tumor specimens in paired comparison with the samples of adjacent non-tumor tissue or normal liver tissue of control patients. In contrast, expression of OX40L mRNA was lower in tumor tissue in paired comparison with the samples of adjacent non-tumor tissue or normal liver tissue. The clinical and pathological analysis showed that expression of OX40 mRNA significantly correlated with the degree of tumor differentiation; there was an insignificant decreasing trend in the length of recurrence-free period. It was hypothesized that microenvironment of hepatocellular carcinoma can induce immunosuppression due to dysregulation of the expression of OX40 and OX40L in tumor tissue, which promotes tumor growth.

[Analysis on clinical characteristics, diagnosis and treatment of 13 cases of infant hairy polyp].

Objective: To explore and analyze the clinical characteristics, diagnosis and treatment of infant hairy polyp. Methods: A retrospective analysis was made on 13 cases of hairy polyp confirmed by pathology, who were admitted to the Children's Hospital of Hebei Province from January 2010 to September 2019, including 4 males and 9 females, with a male-female ratio of 1∶2.25. The age ranged from 3 hours to 1 year, and the median age was 1 month. Twelve of the 13 children were found to have difficulty breathing or feeding. All the children received coblation resection under general anesthesia. The root pedicle of the mass was found in the lateral nasopharyngeal wall in 8 cases, in the junction of palatine and palatopharyngeal arch of tonsil and the tongue and esophageal entrance in 1 case, respectively. Nasal septum was found in 2 cases, including 1 case located between two incisors. The wound at the root pedicle was ablated and the bleeding was stopped completely. Results: Postoperative follow-up lasted from 3 months to 2 years, and there was no recurrence in 12 cases. Fibrolaryngoscope showed a mass of the right eustachian tube and pharyngeal mouth in 1 case 2 years after the surgery, which was considered recurrence of hairy polyps and lost after that. Conclusion: Hairy polyps in infants is a rare clinical disease, and its main symptom is upper respiratory tract obstruction. Early diagnosis and radical surgery are the key to the treatment of the disease.

[Consistency of ALK Ventana-D5F3 immunohistochemistry interpretation in lung adenocarcinoma among Chinese histopathologists].

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer's protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion: There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Structure and mode of action of a novel antibacterial peptide from the blood of Andrias davidianus.

Andrias davidianus is widely recognized in traditional medicine as a cure-all to treat a plethora of ailments. In a previous study, a novel antibacterial peptide named andricin B was isolated from A. davidianus blood. In this study, we investigated andricin B structure and its mode of action. Circular dichroism spectra suggested that andricin B adopts a random coil state in aqueous solution and a more rigid conformation in the presence of bacteria. Moreover propidium iodide/fluorescein diacetate double staining indicated that bacteria treated with andricin B were not immediately eliminated. Rather, there is a gradual bacterial death, followed by a sublethal stage. Scanning electronic microscope imaging indicates that andricin B might form pores on cell membranes, leading to the release of cytoplasmic contents. These results were consistent with flow cytometry analysis. Furthermore, Fourier transform infrared spectroscopy suggests that andricin B induces changes in the chemical properties in the areas surrounding these "pores" on the cell membranes. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study suggested the new perspectives about the mode of action of antimicrobial peptide (AMP) active against sensitive bacteria. The AMP was able to be in a random coiled state in aqueous solution but to change to a more rigid one in the presence of sensitive bacteria. Exposure to AMP might not lead to immediate death of treated bacteria, rather bacteria concentration decreased gradually flattening at a sublethal stage. These findings will help people to understand better how the AMPs activate against sensitive bacteria.

[A novel nanoparticle in treatment of staphylococcus aureus and pseudomonas aeruginosa biofilms].

Objective:To investigate CPC-nanoparticles of low concentrations in treatment of staphylococcus aureus and pseudomonas aeruginosa biofilms in vitro. Method: We established specific biofilms of staphylococcus aureus ATCC 25923 and pseudomonas aeruginosa ATCC 15692, and prepared CPC-nanoparticles and CPC micelle solutions of low concentrations(0.010%, 0.025% and 0.050%). AlamarBlue was used to test the viability of both planktonic staphylococcus aureus and pseudomonas aeruginosa and their biofilms after treatment for 5 minutes and 2 hours respectively in the bactericidal efficacy study.The interaction between CPC-nanoparticles and staphylococcus aureus and pseudomonas aeruginosa biofilms was observed by confocal laser scanning microscope(CLSM). Result: 0.010%, 0.025% and 0.050% CPC-nanoparticles and CPC-micelle solutions had significant bactericidal effect on planktonic staphylococcus aureus and pseudomonas aeruginosa after fiveminute exposure(P<0.05), and staphylococcus aureus and pseudomonas aeruginosa biofilms after both five-minute and two-hour treatments(P<0.05). In CLSM study, the size of staphylococcus aureus biofilms decreased, while dead bacteria of pseudomonas aeruginosa biofilms increased after two-hour treatment. Conclusion: CPC-nanoparticles had significant bactericidal effects on staphylococcus aureus and pseudomonas aeruginosa biofilms, which could be used in treatment of CRS.

Dexmedetomidine enhances ropivacaine-induced sciatic nerve injury in diabetic rats.

BACKGROUND: Previous studies suggest that dexmedetomidine has a protective effect against local anaesthetic-induced nerve injury in regional nerve blocks. Whether this potentially protective effect exists in the context of diabetes mellitus is unknown. METHODS: A diabetic state was established in adult male Sprague-Dawley rats with intraperitoneal injection of streptozotocin. Injections of ropivacaine 0.5%, dexmedetomidine 20 µg kg-1 (alone and in combination), or normal saline (all in 0.2 ml) were made around the sciatic nerve in control and diabetic rats (n=8 per group). The duration of sensory and motor nerve block and the motor nerve conduction velocity (MNCV) were determined. Sciatic nerves were harvested at post-injection day 7 and assessed with light and electron microscopy or used for pro-inflammatory cytokine measurements. RESULTS: Ropivacaine and dexmedetomidine alone or in combination did not produce nerve fibre damage in control non-diabetic rats. In diabetic rats, ropivacaine induced significant nerve fibre damage, which was enhanced by dexmedetomidine. This manifested with slowed MNCV, decreased axon density, and decreased ratio of inner to outer diameter of the myelin sheath (G ratio). Demyelination, axon disappearance, and empty vacuoles were also found using electron microscopy. An associated increase in nerve interleukin-1ß and tumour necrosis factor-α was also seen. CONCLUSIONS: Ropivacaine 0.5% causes significant sciatic nerve injury in diabetic rats that is greatly potentiated by high-dose dexmedetomidine. Although the dose of dexmedetomidine used in this study is considerably higher than that used in clinical practice, our data suggest that further studies to assess ropivacaine (alone and in combination with dexmedetomidine) use for peripheral nerve blockade in diabetic patients are warranted.

SOX2 gene expression and its role in triple negative breast cancer tissues.

The aim of this work was to study the expression of SOX2 gene in triple negative breast cancer and its role. One hundred and twenty specimens of paraffin-embedded triple negative breast cancer (TNBC) tissues were collected from Harbin Medical University Cancer Hospital, Heilongjiang, China between January 2014 and March 2018. The expression of SOX2 was detected using immunohistochemistry, and the relationship between the expression of SOX2 and clinical features was analyzed. Breast cancer cell lines (normal group, SOX2 interference group, SOX2 overexpression group) were cultured in vitro to detect the proliferation and cloning ability of the cell lines. The expression of SOX2 was related to lymph node metastasis and stage of breast cancer (P less than 0.05), but was not related to age, menopause or tumor size (P > 0.05); the expression of SOX2 in the overexpression group was significantly greater than that in the normal group after 72 hours, and no significant difference between the overexpression group and the interference group was observed. The number of clone cells with a diameter of 0.5 mm in the interference group was lower compared to the normal group, and that of the overexpression group was higher, but not significant. SOX2 is associated with the high invasiveness of breast cancer and can be used as a therapeutic target to inhibit the metastasis of cancer cells. SOX2 can promote the proliferation of breast cancer cells and affect the size of clone cells in its involvement in clone.

[Initial study of transthoracic echocardiography guided three-dimensional printing on the application of assessment of structural heart disease].

Objective: To investigate the feasibility and diagnostic value of preoperative transthoracic echocardiography guided three dimensional printing model (TTE Guided 3DPM) on the assessment of structural heart disease (SHD). Methods: From February 2016 to October 2016, 44 patients underwent cardiac surgery in Tianjin Chest Hospital, forty-four patients were assessed preoperatively using TTE Guided 3DPM, including 25 males and 19 females, aged 3-75 years, with an average of (44±22) years. compared to conventional three dimensional transthoracic echocardiography (3D-TTE), and took direct intraoperative findings as "Golden Standard" simultaneously. There are twelve patients with SHD, including four cases with mitral prolapse, two cases with partial endocardial cushion defect, two cases with secondary atrial septal defect, two cases with rheumatic mitral stenosis, one case with tetralogy of Fallot, one case with ventricular septal defect (VSD), thirty-two patients without SHD were designed as negative control. Results: The sensitivity and specificity of TTE Guided 3DPM were greater than or equal to 3D-TTE, P value of McNemar test of 3D-TTE was greater than 0.05, the difference was not statistically significant, kappa=0.745, P<0.01, indicated that the results of 3DTTE and the gold standard were generally consistent.P value of McNemar test of TTE Guided 3DPM was greater than 0.05, the difference was not statistically significant, kappa=0.955, P<0.01, indicated that the results of TTE Guided 3DPM and gold standards were consistent. Compared with 3D-TTE and TTE Guided 3DPM, P value was greater than 0.05, the difference was not statistically significant, kappa=0.879, P<0.01, indicated that the results of 3D-TTE and TTE Guided 3DPM were consistent. TTE Guided 3DPM displayed the three-dimensional structure of SHD cardiac lesions clearly, which were consistent with intraoperative findings. Conclusion: TTE Guided 3DPM provides essential information for the preoperative evaluation and decision of SHD.

Decreased REG1α expression suppresses growth, invasion and angiogenesis of bladder cancer.

BACKGROUND: Previous study has indicated association between REG1α and bladder cancer. The aim of this study was to investigate the role of Regenerating gene I alpha (REG1α) in bladder cancer. METHODS: The role of REG1α in bladder cancer cell proliferation, migration and VEGF-induced angiogenesis was explored in vitro and in vivo. Immunohistochemistry (IHC) analysis was assessed to determine the expression of REG1α in ten paired bladder cancer and adjacent non-cancerous tissues, and in 296 bladder cancer samples. RESULTS: Down-regulation of REG1α expression significantly reduced the proliferation, migration, invasion and VEGF-induced angiogenesis in vitro and the growth of xenograft tumors in vivo. VEGF expression in bladder cancer is associated with REG1α expression and recurrence. REG1α was overexpressed in bladder cancer tissues compared with adjacent normal samples. Patients with elevated REG1α exhibited shorter recurrence times and poor survival. CONCLUSION: Downregulation of REG1α expression can reduce tumor growth, migration, invasion and angiogenesis. Our study demonstrates that REG1α can be used as a marker of recurrence and prognosis in bladder cancer. Therefore, REG1α targeting in bladder cancer patients represents a promising therapeutic strategy.

High expression of KIF14 is associated with poor prognosis in patients with epithelial ovarian cancer.

OBJECTIVE: Kinesin family member 14 (KIF14) is a mitotic kinesin and plays an important role in tumor progression. KIF14 overexpression has been observed in multiple cancers and has been correlated with a poor prognosis. However, its protein expression and prognostic significance in epithelial ovarian cancer (EOC) remain unclear. In this research, we aimed to explore the relationship of KIF14 expression with clinicopathological parameters and prognosis in EOC. MATERIALS AND METHODS: In this study, we measured KIF14 expression in 170 EOC carcinoma tissue samples with immunohistochemistry and correlated these data with clinicopathological characteristics. RESULTS: The expression of KIF14 in EOC tissues was significantly higher than that in normal tissues. Furthermore, KIF14 expression was significantly associated with metastasis (p = 0.047), histological type (p = 0.001), Ki67 expression (p = 0.004) and residual tumor (p = 0.038). Also, Kaplan-Meir survival curves showed that a high level of KIF14 expression was a predictor for worse PFS (p = 0.013) and OS (p = 0.009) in patients with EOC. CONCLUSIONS: KIF14 expression may be associated with poor prognosis, suggesting that it has potential value as an effective prognostic predictor in EOC patients.

Beneficial effects of renal denervation on cardiac angiogenesis in rats with prolonged pressure overload.

AIM: Renal denervation (RDN) has beneficial effects on cardiac remodelling and function in resistant hypertension. We aimed to investigate the impact of RDN on cardiac angiogenesis during prolonged pressure overload. METHODS: Cardiac pressure overload was reproduced by transverse aorta constriction (TAC) procedure in adult Sprague Dawley male rats (n = 35). RDN/sham-RDN procedure was performed in surviving rats at 5 weeks after TAC. RESULTS: Five weeks post-TAC, transthoracic echocardiography revealed that myocardial hypertrophy occurred in TAC rats, with ejection fraction and fractional shortening not significantly changed. At the end of 10 weeks, cardiac systolic function was preserved in RDN group, but not in sham group. CD31 immunohistochemical staining showed that RDN-treated rats had higher cardiac capillary density than sham rats. However, no significant between-group difference was observed in the kidneys. A decreased protein expression of left ventricle vascular endothelial growth factor (VEGF) was observed in sham group, while RDN attenuated this decrease. Compared with sham, RDN resulted in a higher protein expression of VEGF receptor 2 (VEGFR2) and phosphorylated endothelial nitric oxide synthase (p-eNOS) in the heart. CONCLUSION: Renal denervation benefits cardiac angiogenesis during sustained pressure overload, involving regulation of VEGF and VEGFR2 expression as well as activation of eNOS.

Treatment of vitiligo with 308-nm excimer laser: our experience from a 2-year follow-up of 979 Chinese patients.

BACKGROUND: The 308-nm excimer laser had been proved to be a time-efficient and potent therapeutic alternative for the management of vitiligo. Different results had been reported in different ethnic populations. OBJECTIVE: To investigate the efficacy and related contributing factors of 308-nm excimer laser in Chinese vitiligo patients. METHODS: A total of 979 Chinese patients (3478 lesions) with progressive-stage vitiligo who had received 308-nm excimer laser treatment were recruited from the vitiligo clinic of Shandong Provincial Hospital of Dermatology &Venereology from 2012 to 2014. Efficacy of treatment was evaluated at the end of session by two independent dermatologists based on the before and after images taken. Repigmentation was graded on a 4-point scale: grade 1, poor repigmentation (0-25%); grade 2, moderate repigmentation (26-50%); grade 3, good repigmentation (51-75%); grade 4, excellent repigmentation (76-100%). RESULTS: The mean grade of repigmentation was 2.29, 44.22% showed less than 25% repigmentation, 16.27% showed 26-50% repigmentation, 5.95% showed 51-75% repigmentation and 33.55% showed more than 76% repigmentation. The repigmentation of facial lesions was better than lesions located elsewhere (P < 0.0001), the best response was noted in the periorbital region, while lesions on hands and feet showed poor repigmentation (P < 0.0001). The degree of repigmentation was negatively correlated with disease duration (r = -0.268, P < 0.001), age (r = -0.095, P < 0.001) and shape of lesions (r = -0.114, P < 0.001), whereas it was positively correlated with treatment frequency (r = 0.270, P < 0.001). Lesions with concurrent poliosis were more likely resistant to treatments. CONCLUSION: 308-nm excimer laser appears to be an effective and safe treatment in Chinese vitiligo patients. The clinical response and treatment efficacy was affected by many factors such as age, affected anatomical area, shape of the lesion, disease duration and treatment frequencies.