T2-FLAIR mismatch sign and machine learning-based multiparametric MRI radiomics in predicting IDH mutant 1p/19q non-co-deleted diffuse lower-grade gliomas.

AIM: To investigate the application of the T2-weighted (T2)-fluid-attenuated inversion recovery (FLAIR) mismatch sign and machine learning-based multiparametric magnetic resonance imaging (MRI) radiomics in predicting 1p/19q non-co-deletion of lower-grade gliomas (LGGs). MATERIALS AND METHODS: One hundred and forty-six patients, who had pathologically confirmed isocitrate dehydrogenase (IDH) mutant LGGs were assigned randomly to the training cohort (n=102) and the testing cohort (n=44) at a ratio of 7:3. The T2-FLAIR mismatch sign and conventional MRI features were evaluated. Radiomics features extracted from T1-weighted imaging (T1WI), T2-weighted imaging (T2WI), FLAIR, apparent diffusion coefficient (ADC), and contrast-enhanced T1WI images (CE-T1WI). The models that displayed the best performance of each sequence were selected, and their predicted values as well as the T2-FLAIR mismatch sign data were collected to establish a final stacking model. Receiver operating characteristic curve (ROC) analyses and area under the curve (AUC) values were applied to evaluate and compare the performance of the models. RESULTS: The T2-FLAIR mismatch sign was more common in the IDH mutant 1p/19q non-co-deleted group (p<0.05) and the area under the curve (AUC) value was 0.692 with sensitivity 0.397, specificity 0.987, and accuracy 0.712, respectively. The stacking model showed a favourable performance with an AUC of 0.925 and accuracy of 0.882 in the training cohort and an AUC of 0.886 and accuracy of 0.864 in the testing cohort. CONCLUSION: The stacking model based on multiparametric MRI can serve as a supplementary tool for pathological diagnosis, offering valuable guidance for clinical practice.

Prognostic value of multiphase CT angiography: estimated infarct core volume in the patients with acute ischaemic stroke after mechanical thrombectomy.

BACKGROUND AND PURPOSE: Recent studies reported the feasibility of quantifying a reliable infarct core (IC) volume using multiphase computed tomography (mCTA) based on deep learning, however its prognostic value was not fully clarified. Therefore, we aimed to evaluate the prognostic value of mCTA-estimated IC volume in patients with acute ischemic stroke (AIS) after mechanical thrombectomy (MT). MATERIALS AND METHODS: We retrospectively reviewed patients who underwent mCTA and MT for large vessel occlusion in middle cerebral artery and (or) internal carotid artery within 6 hours after symptom onset between January 2018 and November 2019. Patients were dichotomized into good (modified Rankin Scale [mRS] score, 0-2) and poor (mRS, 3-6) outcome groups. mCTA-estimated IC volume were generated based on a multi-scale three-dimensional convolutional neural network. Univariate, multivariate logistic regression and receiver operating characteristic (ROC) curve analyses were used to identify the independent variables, and evaluate their performances in predicting the clinical outcome. RESULTS: Of 44 included patients, 27 (61.4%) patients achieved good outcome. National Institutes of Health Stroke Scale scores at admission [NIHSSpre] (odds ratio [OR], 1.191; 95%confidence interval [CI], 1.028-1.379; P=0.020) and mCTA-estimated IC volume (OR, 1.076; 95%CI, 1.016-1.140; P=0.013) were found to be independently associated with functional outcome in patients with AIS after MT. After integrating NIHSSpre and mCTA-estimated IC volume, optimal performance (area under the ROC curve, 0.874; 95%CI, 0.739-0.954) could be obtained in predicting the clinical outcome. CONCLUSIONS: mCTA-estimated IC volume might be promising for predicting the prognosis, and assisting in making individualized treatment decision in patients with AIS.

[Nodal T-follicular helper cell lymphoma, angioimmunoblastic-type associated with diffuse large B-cell lymphoma: a clinicopathological study].

Objective: To investigate the clinicopathological features and molecular genetics of diffuse large B-cell lymphomas (DLBCL) with concurrent or secondary to nodal T-follicular helper cell lymphoma, angioimmunoblastic-type (nTFHL-AI). Methods: The clinicopathological features and molecular genetics of DLBCL associated with nTFHL-AI diagnosed between January 2015 and October 2022 at the First Affiliated Hospital of Zhengzhou University were analyzed using histology, immunohistochemistry, PCR, EBV-encoded RNA in situ hybridization and fluorescence in situ hybridization (FISH). Clinical information was collected and analyzed. Results: A total of 6 cases including 3 nTFHL-AI with secondary DLBCL and 3 composite lymphomas were reviewed. There were 4 male and 2 female patients, whose ages ranged from 40 to 74 years (median 57 years). All patients presented with nodal lesions at an advanced Ann Arbor stage Ⅲ/Ⅳ (6/6). Bone marrow involvement was detected in 4 patients. All cases showed typical histologic and immunophenotypic characteristics of nTFHL-AI. Among them, 5 cases of DLBCL with concurrent nTFHL-AI exhibited numerous large atypical lymphoid cells and the tumor cells were CD20 and CD79α positive. The only case of DLBCL secondary to nTFHL-AI showed plasma cell differentiation and reduced expression of CD20. All of cases were activated B-cell (ABC)/non-germinal center B-cell (non-GCB) subtype. Three of the 6 cases were EBV positive with>100 positive cells/high power field, meeting the diagnostic criteria of EBV+DLBCL. The expression of MYC and CD30 protein in the DLBCL region was higher than that in the nTFHL-AI region (n=5). C-MYC, bcl-6 and bcl-2 translocations were not detected in the 4 cases that were subject to FISH. Four of the 6 patients received chemotherapy after diagnosis. For the DLBCL cases of nTFHL-AI with secondary DLBCL, the interval was between 2-20 months. During the follow-up period ranging from 3-29 months, 3 of the 6 patients died of the disease. Conclusions: DLBCL associated with nTFHL-AI is very rare. The expansion of EBV-infected B cells in nTFHL-AI may progress to secondary EBV+DLBCL. However, EBV-negative cases have also been reported, suggesting possible other mechanisms. The up-regulation of MYC expression in these cases suggests a possible role in B-cell lymphomagenesis. Clinicians should be aware that another biopsy is still necessary to rule out concurrent or secondary DLBCL when nodal and extranodal lesions are noted after nTFHL-AI treatment.

Hypoperfusion intensity ratio correlates with clinical outcome of endovascular thrombectomy in acute ischaemic stroke patients with late therapeutic window.

AIM: To evaluate the prognostic value of the hypoperfusion intensity ratio (HIR) on 90-day clinical outcome in acute ischaemic stroke (AIS) patients with late therapeutic window. MATERIALS AND METHODS: One hundred and sixty-eight consecutive AIS patients with anterior-circulation large-vessel occlusion who underwent endovascular thrombectomy during the late window were enrolled retrospectively. Clinical data, Alberta Stroke Program Early Computed Tomography Score (ASPECTS) based on unenhanced computed tomography (CT), and perfusion parameters included ischaemic core, hypoperfusion volume, mismatch volume between core and penumbra, and the HIR were assessed and compared between patients with or without favourable outcomes (defined as modified Rankin Scale score of 0-2). Statistical analysis included binary logistic regression and receiver operating characteristic (ROC) analyses. RESULTS: A favourable outcome was achieved in 76 (45.2%) patients. In univariable analysis, age, National Institutes of Health Stroke Scale (NIHSS) score at admission, ASPECTS score, HIR, ischaemic core, and hypoperfusion volume were significantly associated with functional outcome (p<0.05). In multivariate analyses, age (OR 0.95; 95% CI 0.92-0.99), NIHSS score at admission (OR 0.89, 95% CI 0.84-0.96) and HIR (OR 0.018, 95% CI 0.003-0.113) remained as independent outcome predictors (p<0.01). The optimal threshold of HIR was 0.36 (sensitivity 70.7%, specificity 61.8%). The combination of age, NIHSS score at admission, and HIR yield good performance for outcome prediction with an area under the ROC curve of 0.815 (sensitivity 88.2%, specificity 64.1%), significantly higher than individual variable (p<0.05). CONCLUSION: Low HIR was a predictor for favourable outcome in AIS patients with late therapeutic window. Integrating HIR with clinical variables improved the ability for outcome classification.

[Effect of autologous hematopoietic stem cell transplantation on minimal residual disease in patients with multiple myeloma].

Objective: To evaluate the effect of autologous hematopoietic stem cell transplantation (ASCT) on minimal residual disease (MRD) in patients with multiple myeloma (MM). Method: From August 2018 to August 2021, 92 patients newly diagnosed with MM who had received either the bortezomib combined with cyclophosphamide and dexamethasone (VCD) or the bortezomib, lenalidomide and dexamethasone (VRD) induction regimens followed by sequential ASCT were assessed for overall survival (OS) and the MRD negative rate. The differences in efficacy at 100 days after transplantation were assessed according to factors, including age, risk stratification, target organ damage, and pre-transplant regimen, etc. Results: Among the 92 patients, there were 45 males and 47 females, with a median age of 57.3 (35-67) years. Fifty-seven patients received the VCD regimen, and 35 received VRD as induction regimen. Forty-three patients received busulphan combined with cyclophosphamide and etoposide (BCV), and 49 patients received high-dose melphan (HDM) regimen as pre-transplantation treatment. After transplantation, the total complete remission (CR) rate of 92 patients increased from 23.9% (22/92) to 58.7% (54/92), and the MRD negative rate increased from 4.4% (4/92) to 33.7% (31/92), and the differences were statistically significant (all P<0.05). After transplantation, the MRD negative rates of patients with PR, VGPR and ≥CR before transplantation were 17.6% (6/34), 33.3% (12/36) and 59.1% (13/22), respectively (P=0.006). The CR rates of patients with or without plasmacytoma at initial diagnosis were 36.4% (4/11) and 65.4% (53/81), respectively (P=0.029), and the MRD negative rates were 18.2% (2/11) and 39.5% (32/81), respectively (P=0.037), and the differences were statistically significant. The MRD negative rates in high-risk patients and standard-risk group were 30.5% (12/28) and 42.9% (18/59), respectively (P=0.258). For patients who achieved efficacy above VGPR before transplantation, the MRD negative rates after transplantation in VCD-induced group and VRD group were 29% (9/31) and 59.3% (16/27), respectively (P=0.033), and in BCV group and HDM group were 24% (6/25) and 57.6% (19/33), respectively (P=0.016), the differences between the groups were both statistically significant. Conclusion: ASCT can overcome the adverse factors such as high-risk cytogenetic abnormalities, and significantly improve the CR rate and MRD negative rate of MM patients. However, the benefit for patients with plasmacytoma at initial diagnosis is not as good as that of patients without.

[Characteristics of liver function changes in 111 elderly patients with COVID-19 pneumonia].

Objective: To retrospectively analyze the characteristics and influencing factors of liver function changes in 111 elderly patients with COVID-19 pneumonia. Methods: 111 elderly patients with COVID-19 admitted to the Department of Infectious Diseases, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology from February 5 to March 3, 2020 were enrolled. According to the severity of disease and liver function condition, they were divided into severe group (n=40), normal group (n=71), abnormal liver function group (n=86) and normal liver function group (n=25). The indexes related to liver function changes [total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT)] and related influencing factors were analyzed. Results: Among 111 cases, 86 (77.5%) had abnormal liver function of varying degrees, and 28 (25.2%) had liver injury. The abnormal rates of TBil, AST, ALP and GGT were significantly higher in the severe group than normal group (P<0.05). There were no significant differences in age, ribavirin, glucocorticoid and the application of lopinavir-ritonavir tablets between the abnormal liver function and the normal group (P>0.05). The proportion of male was significantly higher in the abnormal liver function than normal liver function group (P<0.05). Conclusion: Elderly COVID-19 patients have a higher proportion of abnormal liver function, and patients in the severe group are more likely to have higher level of TB, AST, ALP and GGT. The abnormal liver function may be related to the direct viral infection of the liver and the inflammatory immune response of the body after infection in elderly patients.

A radiomics-based model to differentiate glioblastoma from solitary brain metastases.

AIM: To differentiate glioblastoma (GBM) from solitary brain metastases (MET) using radiomic analysis. MATERIALS AND METHODS: Two hundred and fifty-three patients with solitary brain tumours (157 GBM and 98 solitary brain MET) were split into a training cohort (n=178) and a validation cohort (n=77) by stratified sampling using computer-generated random numbers at a ratio of 7:3. After feature extraction, minimum redundancy maximum relevance (mRMR) and the least absolute shrinkage and selection operator (LASSO) were used to build the radiomics signature on the training cohort and validation cohort. Performance was assessed by radiomics score (Rad-score), receiver operating characteristic (ROC) curve, calibration, and clinical usefulness. RESULTS: Eleven radiomic features were selected as significant features in the training cohort. The Rad-score was significantly associated with the differentiation between GBM and solitary brain MET (p<0.001) both in the training and validation cohorts. The radiomics signature yielded area under the curve (AUC) values of 0.82 and 0.81 in the training and validation cohorts to distinguish between GBM and solitary brain MET. CONCLUSIONS: The radiomics model might be a useful supporting tool for the preoperative differentiation of GBM from solitary brain MET, which could aid pretreatment decision-making.

Combined texture analysis of diffusion-weighted imaging with conventional MRI for non-invasive assessment of IDH1 mutation in anaplastic gliomas.

AIM: To examine whether texture analysis (TA) of diffusion-weighted imaging (DWI) combined with conventional magnetic resonance imaging (MRI) could non-invasively predict isocitrate dehydrogenase 1 (IDH1) mutational status in anaplastic gliomas. MATERIALS AND METHODS: Fifty-two patients with histologically confirmed anaplastic glioma was reviewed retrospectively. Conventional MRI was evaluated using the Visually Accessible Rembrandt Images (VASARI) scoring system. TA of DWI based on the entire tumour volume was compared between IDH1-mutant and wild-type tumours by using unpaired Student's t-test. Receiver operating characteristic curve (ROC) and logistic regression were used to assess their diagnostic performance. RESULTS: Significant statistical differences in VASARI features and TA of DWI were observed between IDH1-mutant and wild-type tumours (all p<0.05). Using multivariable logistic regression, the proportion of the tumour that was non-enhancing and the entropy of apparent diffusion coefficient (ADC) were found to possess higher prediction potential for IDH1 mutation with areas under the ROC curve (AUC) of 0.918 and 0.724, respectively. A combination of these for the identification of IDH1 mutations improved the AUC to 0.954, with a sensitivity and a specificity of 81% and 96%. CONCLUSIONS: The combined assessment of the conventional MRI and TA of DWI were useful for predicting IDH1 mutation in anaplastic gliomas.

[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.

[Application of uterine lower part breakwater-like suture operation in placenta previa].

Objective: To explore the efficacy and safety of uterine lower posterior wall breakwater-like suture technique in controlling the intraoperative bleeding of placenta previa. Methods: From June 2016 to June 2017, 47 patients were diagnosed placenta previa in Union Hospital, Tongji Medical College of Huazhong University of Science and Technology. Posterior wall breakwater-like suture technique was used preferentially, as for cases with poor myometrium layer, lower anterior wall stitch suture was used at the same time. Bilateral descending branches of uterine artery ligation and Cook balloon compression of uterine lower segment was conducted when necessary. The clinic data of the 47 cases were analyzed. Results: Thirty cases (63.8, 30/47) were diagnosed placenta inccreta or percreta by ultrasound or MRI preoperatively. Senventeen cases were diagnosed as placenta accreta (36.2%, 17/47) . Thirty-four cases had the previous history of cesarean section. The average cervical canal length of 47 patients was (2.8±0.9) cm. There were 19 cases (40.4%,19/47) with 1 time posterior wall breakwater-like sutured and 16 cases (34.0%,16/47) with 2 or 3 times posterior wall breakwater-like sutured; 12 cases (25.5%,12/47) were treated with anterior wall stitch suture simultaneously.Ten cases (21.3%, 10/47) underwent uterine artery ligation, 17 cases (36.2%, 17/47) underwent COOK balloon compression on the staxis surface of lower segment. None of them had postpartum hemorrhage or performed internal iliac artery embolization. The median blood loss in the operation was 700 ml, the percentiles 25 was 500 ml, and the percentiles 75 was 1 200 ml. The blood loss ≥1 000 ml in 18 (38.3%, 18/47) patients,and the most serious one was 2 500 ml. The median blood transfusion volume (including allogenetic transfusion and autotransfusion) was 450 ml, the percentiles 25 was 228 ml, and the percentiles 75 was 675 ml. The average vaginal bleeding volume was (150±63) ml first day after operation. The mean hospitalization time was (4.7±1.0) days. The mean gestational weeks of pregnancy termination was (36.1±1.5) weeks, and the mean birth weight of newborns was (2 817±492) g. Apgar score:1-minute 7.8±1.1, 5-minute 8.9±0.8. No neonatal death, 16 cases were transferred to neonatal ICU (34.0%, 16/47) mainly for premature delivery and low birth weight. No complication was found in 6 months post-operation. Conclusions: Uterine posterior wall breakwater-like suture technique is a simple, safe and effective way in controlling intraoperative bleeding of placental previa.Lower anterior wall stitch suture could effectively stop bleeding and restore the normal uterine shape. Combined application of various methods could significantly reduce the incidence of postpartum hemorrhage and hysterectomy, and improve maternal and fetal prognosis.

Quantitative evaluation of diffusion-kurtosis imaging for grading endometrial carcinoma: a comparative study with diffusion-weighted imaging.

AIM: To evaluate the diagnostic performance of histogram analysis of diffusion kurtosis magnetic resonance imaging (DKI) and standard diffusion-weighted imaging (DWI) in discriminating tumour grades of endometrial carcinoma (EC). MATERIALS AND METHODS: Seventy-three patients with EC were included in this study. The apparent diffusion coefficient (ADC) value from standard DWI, apparent diffusion for Gaussian distribution (Dapp), and apparent kurtosis coefficient (Kapp) from DKI were acquired using a 3 T magnetic resonance imaging (MRI) system. The measurement was based on an entire-tumour analysis. Histogram parameters (Dapp, Kapp, and ADC) were compared between high-grade (grade 3) and low-grade (grade 1 and 2) tumours. The diagnostic performance of imaging parameters for discriminating high- from low-grade tumours was analysed using a receiver operating characteristic curve (ROC). RESULTS: The area under the ROC curve (AUC) of the 10th percentile of Dapp, 90th percentile of Kapp and 10th percentile of ADC were higher than other parameters in distinguishing high-grade tumours from low-grade tumours (AUC=0.821, 0.891 and 0.801, respectively). The combination of 10th percentile of Dapp and 90th percentile of Kapp improved the AUC to 0.901, which was significantly higher than that of the 10th percentile of ADC (0.810, p=0.0314) in differentiating high- from low-grade EC. CONCLUSION: Entire-tumour volume histogram analysis of DKI and standard DWI were feasible for discriminating histological tumour grades of EC. DKI was relatively better than DWI in distinguishing high-grade from low-grade tumour in EC.

The Cold Hardiness of Phrynocephalus erythrurus, the Lizard Living at Highest Altitude in the World.

BACKGROUND: Phrynocephalus erythrurus living at Qinghai-Tibet Plateau, is believed to be the highest lizard in the world, but we know little about how these lizards cope with very low temperatures in winter. OBJECTIVE: The aim of this study was to find the difference of the lizards before and after cold acclimatization. MATERIALS AND METHODS: In this study the limit of supercooling and inoculative freezing, the concentration of four organic osmolytes, and the activity of lactate dehydrogenase in the plasma were measured in samples shortly after capture and in other samples after 7~8 weeks of acclimatization at 2~4 degree C. RESULTS: Animals acquired an ability to undergo deeper supercooling and inoculative freezing through the course of acclimatization. We find no regular changes of the four organic osmolytes after the acclimatization. CONCLUSION: We think that this species of lizard is partly freeze-tolerant and conclude that it uses supercooling to survive in winter.

Comparison of Mitochondrial Function in Boar and Bull Spermatozoa Throughout Cryopreservation Based on JC-1 Staining.

BACKGROUND: Poor reproductivity hampers the commercialization of cryopreserved boar semen. OBJECTIVE: This study was to determine the differences in the sperm mitochondrial function between boar and bull semen at different cryopreservation stages. MATERIALS AND METHODS: Boar and bull fresh, equilibrated, and frozen-thawed spermatozoa were evaluated for mitochondrial function using JC-1 under a fluorescent microscope. RESULTS: Bull and boar percentage of spermatozoa staining green (PSSG) showed no difference between fresh and equilibrated semen (P> 0.05). However, frozen-thawed bull and boar semen demonstrated significantly higher PSSG (P < 0.01) than fresh and equilibrated semen. Frozen-thawed boar semen represented a significantly higher PSSG (P < 0.01) than bull semen. CONCLUSION: Negative cryopreservation influence on boar and bull spermatozoa was not significantly produced by pre-freezing procedures, but rather by freezing and thawing. Cryopreservation has more pronounced negative effects on boar than on bull spermatozoa, which partly explains lagged commercialization of frozen boar semen.

[Fluorescence in situ hybridization combined with cytomorphology for the detection of lung cancer in bronchial brushing specimens].

Objective: To evaluate the diagnostic value of fluorescence in situ hybridization (FISH) combined with bronchial brushing cytology for detecting lung cancer. Methods: Centromeric enumeration probes (CEPs) for chromosomes 7, 8 and 17 were used in FISH assay. The combination of FISH and cytology was analyzed in 69 bronchial brushing specimens. Results: The positive rates of CEP7, CEP8 and CEP17 in malignant cases diagnosed by cytology were 50.0%, 80.8% and 65.4%, respectively. CEP8 probe showed significantly higher positive rate than CEP7 (P=0.015). In the samples of suspicious of malignancy, the positive rates of CEP7, CEP8 and CEP17 were 46.6%, 66.7% and 58.8%, respectively. While in atypical cases, the positive rates of these three probes were 20.0%, 33.3% and 25.0%, respectively. There was no statistical difference between suspicious of malignancy and atypical cases (P>0.05) as well as between malignant and suspicious of malignancy (P>0.05). No chromosome aberrations were found in normal cases diagnosed by cytology. The positive rates of these three probes in adenocarcinoma (ADC) were slightly higher than those in squamous cell carcinoma and small cell lung cancer. However, only CEP8 probe showed statistically difference between ADC and small cell lung cancer (P=0.044). The combination of cytology and FISH using any one of the three-probe set (CEP7, CEP8 and CEP17) showed the sensitivity and specificity of 80.3% and 100.0%, while those of cytology were 54.1% and 100.0%, respectively. Conclusions: FISH combined with cytomorphology assisted the cytology diagnosis of suspicious of malignancy and atypical cases. Therefore, it significantly improved the diagnostic sensitivity for lung cancer without sacrificing specificity.

The significance of miR-145 in the prediction of preeclampsia.

AIM: The aim of this study was to explain the effects of miRNA-145 in the pathogenesis of preeclampsia. METHODS: Collecting the placental tissue of 40 severe preeclampsia patients and 20 normal pregnant women, and observation of the pathological findings by HE staining. Measuring the miR-145 by RT-PCR. EVCT were divided into NC group; MC group and miRNA group. The EVCT cells of MC and miRNA groups were simulated by hypoxia in vivo by CoCl2. Measuring the proliferation rate of different groups by MTT testing. The cells apoptosis rates were measured by flow cytometry; evaluating PI3K, Akt, mTOR and P53 gene and protein expression of three groups by RT-PCR and WB. RESULTS: Compared to the normal pregnant placental tissue. The miR-145 expression of preeclampsia pregnant placental tissue was significantly decreased (p < 0.05). In the cell experiments, the proliferation rate was significantly increased, and the cell apoptosis rate was significantly reduced in MC group compared to the MC group (p<0.05, respectively). Comparing with MC group, the PI3K, Akt and mTOR gene and protein expression of miRNA group were significantly up-regulated and the P53 expression was significantly down-regulated (p<0.05, respectively). CONCLUSION: miR-145 might have effects to predict preeclampsia via PI3K/Akt/mTOR signalling pathways (Fig. 5, Ref. 30).

Stability of the cytoskeleton of matured buffalo oocytes pretreated with cytochalasin B prior to vitrification.

Stabilizing the cytoskeleton system during vitrification can improve the post-thaw survival and development of vitrified oocytes. The cytoskeleton stabilizer cytochalasin B (CB) has been used in cryopreservation to improve the developmental competence of vitrified oocytes. To assess the effect of pretreating matured buffalo oocytes with CB before vitrification, we applied 0, 4, 8, or 12 µg/mL CB for 30 min. The optimum concentration of CB treatment (8 µg/mL for 30 min) was then used to evaluate the distribution of microtubules and microfilaments, the expression of the cytoskeleton proteins actin and tubulin, and the developmental potential of matured oocytes that were vitrified-warmed by the Cryotop method. Western blotting demonstrated that vitrification significantly decreased tubulin expression, but that the decrease was attenuated for oocytes pretreated with 8 µg/mL CB before vitrification. After warming and intracytoplasmic sperm injection, oocytes that were pretreated with 8 µg/mL CB before vitrification yielded significantly higher 8-cell and blastocyst rates than those that were vitrified without CB pretreatment. The values for the vitrified groups in all experiments were significantly lower (P < 0.01) than those of the control groups. In conclusion, pretreatment with 8 µg/mL CB for 30 min significantly improves the cytoskeletal structure, expression of tubulin, and development capacity of vitrified matured buffalo oocytes.

Isolation, proliferation, and induction of Bama mini-pig spermatogonial stem cells in vitro.

Spermatogonial stem cells (SSCs), the unique seed cells of testes, can undergo meiosis and form spermatozoa, thus transmitting genetic information to offspring. Research concerning these cells explores the mechanism underlying spermatogenesis, making possible the induction of their differentiation into spermatozoa in vitro. SSCs have therefore attracted much interest among scientists. Although the proliferation of such cells in vitro has been demonstrated, we are unaware of any long-term laboratory culture of porcine SSCs. The objective of this study was to isolate, characterize, culture, and induce the differentiation of Bama mini-pig SSCs. SSCs were isolated using differential plating and cultured for over 100 days on an STO feeder cell layer without serum. Cell clusters appeared after three passages and continuously formed during subsequent cultivation. Staining showed that these clusters were positive for UCHL1 and CDH1, could be bound by Dolichos biflorus agglutinin, and that some cells expressed OCT4. Ultrastructure observations revealed SSCs in testis tissue to be round in shape, while those cultured in vitro were flat and bound together. Our attempts at inducing differentiation showed that SSCs cultured in vitro could undergo meiosis. In this study, we describe an effective culture system for Bama mini-pig SSCs capable of producing enough cells to establish a platform for further SSC research, such as genetic manipulation or exploration of the mechanism underlying spermatogenesis.

Production of hGFAP-DsRed transgenic Guangxi Bama mini-pigs via somatic cell nuclear transfer.

The mini-pig is a useful animal model for human biomedical research due to its physiological similarity to humans and the ease of handling. In order to optimize the efficiency of production of transgenic Bama mini-pigs through somatic cell nuclear transfer (SCNT), we examined the effects of contact inhibition, roscovitine treatment, and serum starvation on the cell cycle synchronization and transgenic cloned embryo development in vivo and in vitro after nuclear transfer. The analysis showed that the rates of G0/G1 stage cells in the contact inhibition (92.11%) and roscovitine treatment groups (89.59%) were significantly higher than in serum starvation group (80.82%). A higher rate of apoptosis was seen in the serum starvation group (14.13%) compared to the contact inhibition and roscovitine treatment groups (6.71 and 2.46% respectively, P < 0.05). There was a significant decrease in blastocyst yield in the serum starvation group (14.19%) compared to the roscovitine treatment and contact inhibition groups (21.31 and 20.32% respectively, P < 0.05). A total of 1070 transgenic cloned embryos derived from the three treatment groups were transferred to surrogate sows; one pregnancy was established and three embryos from the roscovitine treatment group successfully completed gestation. These results indicate that the roscovitine treatment was more effective at synchronizing transgenic kidney cells in Bama mini-pigs and allowed reconstructed embryos to develop to full term.

Endovascular Recanalization of Symptomatic Chronic ICA Occlusion: Procedural Outcomes and Radiologic Predictors.

BACKGROUND AND PURPOSE: Endovascular recanalization has been attempted in patients with symptomatic chronic ICA occlusion, however, the heterogeneity of recanalization outcomes and the perioperative complications present challenges for the clinical application. Our aim was to evaluate the safety and efficacy of endovascular recanalization for symptomatic chronic ICA occlusion and identify potential predictors for successful recanalization. MATERIALS AND METHODS: This study included 47 consecutive patients with symptomatic chronic ICA occlusion who underwent endovascular recanalization at our institution. Patients' clinical information, radiologic characteristics, procedural results, and outcomes were recorded. Factors related to technical success were analyzed by univariate and multivariate analyses. RESULTS: The technical success rate was 74.5% (35/47); 12.8% of patients (6/47) experienced intraoperative complications, but none had permanent neurologic deficits. Three months after recanalization, 21 of the 29 recanalized patients (72.4%) and 3 of the 10 failed patients (30.0%) demonstrated improved mRS scores. Restenosis or re-occlusion occurred in 12.9% of patients (4/31) with successful recanalization. Multivariate analysis showed that tapered or blunt stump (P = .016), distal ICA occlusion segment (below the cavernous segment versus at or above the ophthalmic segment, P = .003; at the cavernous or clinoid segment versus at or above the ophthalmic segment, P = .027), and radiologic occlusion to recanalization of ≤3 months (P = .044) were significantly associated with successful recanalization. Patients were assigned points according to the coefficients of the prediction model, and the technical success rates were 0%, 46.2%, 90.5%, and 100% in patients with 1, 2, 3, and 4 points, respectively. CONCLUSIONS: Endovascular recanalization is a safe and effective treatment for symptomatic chronic ICA occlusion in selected patients. A residual stump, low levels of the distal ICA occlusion segment, and a short radiologic occlusion time were identified as positive predictors of technical success.