[Expression of DARS2 in colorectal cancer and its clinical significance].

Objective: To investigate the expression of DARS2 and its clinical significance in colorectal cancer. Methods: In this study, bioinformatics tools, especially gene expression profile interactive analysis 2 (GEPIA2), were used to conduct an in-depth analysis of DARS2 expression in colorectal cancer tissues. Immunohistochemical staining was carried out in 108 colorectal cancer specimens and 30 normal colorectal tissues obtained from the First Affiliated Hospital of Nanchang University, Nanchang, China. Colorectal cancer cell lines (HCT116 and SW480) were transfected with small interfering RNA (siRNA) and DARS2 overexpression plasmid to examine the effects of DARS2 knockdown and overexpression on cell function. To assess the effects on cell function, CCK8 and transwell migration assays were used to assess proliferation and cell motility, respectively. Additionally, protein immunoblotting was employed to scrutinize the expression of proteins associated with the epithelial-mesenchymal transition of colorectal cancer cells. Results: DARS2 exhibited a pronounced upregulation in expression within colorectal cancer tissues compared to their normal epithelial counterparts. Furthermore, DARS2 expression was higher in colorectal cancer of stage Ⅲ-Ⅳ than those of stage Ⅰ-Ⅱ, exhibiting a significant correlation with N staging, M staging, and pathological staging (P<0.05). Kaplan-Meier analyses showed a decreased overall survival rate in colorectal cancer with DARS2 expression compared to those without DARS2 expression (P<0.05). In the siRNA transfection group, there was a significant reduction in cell proliferation and migration (P<0.01 and P<0.05, respectively). Conversely, the transfection of DARS2 overexpression plasmids substantially increased both cell proliferation and migration (P<0.05). Additionally, immunoblotting revealed that DARS2 knockdown led to an upregulation of E-cadherin expression and a downregulation of N-cadherin and vimentin expression. In contrast, DARS2 overexpression resulted in increased N-cadherin and vimentin expression, coupled with reduction in E-cadherin expression. Conclusions: There is a strong association between DARS2 expression and colorectal cancer progression. Silencing DARS2 inhibits cell proliferation and migration, exerting a discernible influence on the epithelial-mesenchymal transition process.

Deep-learning model associating lateral cervical radiographic features with Cormack-Lehane grade 3 or 4 glottic view.

Unanticipated difficult laryngoscopy is associated with serious airway-related complications. We aimed to develop and test a convolutional neural network-based deep-learning model that uses lateral cervical spine radiographs to predict Cormack-Lehane grade 3 or 4 direct laryngoscopy views of the glottis. We analysed the radiographs of 5939 thyroid surgery patients at our hospital, 253 (4%) of whom had grade 3 or 4 glottic views. We used 10 randomly sampled datasets to train a model. We compared the new model with six similar models (VGG, ResNet, Xception, ResNext, DenseNet and SENet). The Brier score (95%CI) of the new model, 0.023 (0.021-0.025), was lower ('better') than the other models: VGG, 0.034 (0.034-0.035); ResNet, 0.033 (0.033-0.035); Xception, 0.032 (0.031-0.033); ResNext, 0.033 (0.032-0.033); DenseNet, 0.030 (0.029-0.032); SENet, 0.031 (0.029-0.032), all p < 0.001. We calculated mean (95%CI) of the new model for: R2 , 0.428 (0.388-0.468); mean squared error, 0.023 (0.021-0.025); mean absolute error, 0.048 (0.046-0.049); balanced accuracy, 0.713 (0.684-0.742); and area under the receiver operating characteristic curve, 0.965 (0.962-0.969). Radiographic features around the hyoid bone, pharynx and cervical spine were associated with grade 3 and 4 glottic views.

[The relationship between resting heart rate and all-cause mortality among the Chinese oldest-old aged more than 80: a prospective cohort study].

Objective: To explore the association between resting heart rate(RHR) and all-cause mortality among the Chinese oldest-old aged more than 80. Methods: Using a total of seven surveys or follow-ups data (1998, 2000, 2002, 2005, 2008, 2011 and 2014) from the Chinese Longitudinal Healthy Longevity Survey (CLHLS). A total of 17 886 elderly over 80 years old were selected as subjects, their resting heart rate were measured though baseline survey and the survival outcome and death time of the subjects were followed up. The subjects were divided into 6 groups according to their resting heart rate. Cox regression model was used to estimate the effect of resting heart rate on mortality risk. The interaction of age, gender and resting heart rate was also analyzed by likelihood ratio test. Results: The age of subjects M (P25, P75) was 92 (86, 100) years old, including 10 531 females (58.9%) and there were 13 598 participants died, the mortality rate was 195.5 per 1 000 person-years. Multivariate Cox regression analysis showed that compared to the control group (60-69 pbm/min), the hazard ratio of the elderly are 1.06 (95%CI: 1.02, 1.11), 1.09 (95%CI: 1.04, 1.15), 1.23 (95%CI: 1.14, 1.34), 1.25 (95%CI: 1.08, 1.44) in the group of RHR between 70-79, 80-89, 90-99 and ≥100 pbm/min and P values are all less than 0.05. Likelihood ratio test showed that RHR and age had an interaction effect. (P for interaction=0.011). Conclusion: The risk of all-cause death increased with the increase of resting heart rate and this relationship was stronger between the 80-89 years old people.

[The preventive effect of four drugs on acute mountain sickness: a Bayesian network meta-analysis].

Objective: To compare and predict the preventive effects of acetazolamide and other drugs on acute mountain sickness(AMS). Methods: Following the retrieval strategy of PRISMA statement of systematic review and meta-analysis, we searched the databases of PubMed, Embase, Cochrane Library, Web of Science, CNKI, Wanfang, etc. from January 1, 1980 to November 30, 2020, and randomized controlled trials (RCT) consistent with drug prevention of AMS were conducted. Using R and other statistical software, Markov chain-Monte Carlo method was carried out for network meta-analysis under Bayesian framework, and node separation method was performed to check the consistency of closed-loop research. Results: Twenty-three literatures (25 studies) were included to compare the preventive effects of 4 drugs on AMS. Bayesian network meta-analysis showed that the incidence of AMS in acetazolamide group (ACE), dexamethasone group (DEX), ginkgo biloba extract group (GBE) and rhodiola group (RHO) was lower than that in placebo group (PLA). In the comparison of drug groups, the incidence of AMS in ACE, DEX and RHO was lower than that in GBE. There was no statistically significant difference in the incidence of AMS among ACE, DEX and RHO groups. Eight of these studies reported the effects of two drugs on pulse oxygen saturation (SpO2) in people entering the target altitude. Bayesian network meta-analysis showed that SpO2 in RHO was higher than that in ACE and PLA, but there was no statistically significant difference in SpO2 between ACE and PLA. The probability ranking of prevention AMS effect grade showed that the rank 5th probability of AMS in ACE, DEX, GBE, RHO and PLA was 45.72%, 48.80%, 0, 5.48% and 0, respectively. The probability ranking of improving the SpO2 level of the target altitude population showed that the probability of the ACE, RHO and PLA ranking 1st in improving the SpO2 effect at the target altitude was 2.27%, 97.66% and 0.07%, respectively; the results of direct comparison were in good agreement with those of Bayesian prediction model indirectly, and there was no statistical difference. Conclusions: Acetazolamide and dexamethasone can effectively prevent AMS, and should be the first choice for related supplementary research in the future. Rhodiola not only improves the SpO2 of people entering high altitude, but also reduces the incidence of AMS, which needs more attention. Ginkgo biloba extract is not as effective as the above three drugs in preventing AMS and should be used depending on clinical situations.

[Research progress on non-alcoholic fatty liver disease animal models].

In recent years, with the changes in living standards and dietary structure, the incidence of non-alcoholic fatty liver disease has been increasing year by year in China, and the incidence rate in the general population is as high as 29.81%. An increasingly epidemiological evidence suggests that non-alcoholic fatty liver disease has become one of the causes of increasing liver cirrhosis and liver cancer. However, its etiology and pathogenesis are complex and have not yet been fully elucidated. Therefore, establishing an appropriate non-alcoholic fatty liver disease animal models for pre-clinical research is essential to elucidate its pathogenesis. This article summarizes the latest research progress of non-alcoholic fatty liver disease animal models, which are common at home and abroad in recent years.

[Evaluation of different staging systems and prognostic analysis of 110 primary gastrointestinal diffuse large B cell lymphoma].

Objective: To compare the prognostic efficiency of Lugano staging, TNM staging and Musshoff staging systems in patients with primary gastrointestinal diffuse large B-cell lymphoma(PGI-DLBCL) and investigate its clinical features and prognosis. Methods: The clinical data of 110 patients with PGI-DLBCL in Tianjin Medical University Cancer Institute and Hospital from May 2008 to August 2017 was retrospectively analyzed. The stage of lymphoma was assessed following Lugano staging, TNM staging and Musshoff staging systems respectively. The prognostic value was compared mainly according to the situation of 5-year overall survival (OS)and the influence of different clinical features on prognosis of patients was also investigated. Results: The median age of the whole study was 55(range 17-92) years old. With a median follow-up time of 36 (range 1-115) months, the median progression-free survival (PFS) was 35 (range 0-86) months, and the median overall survival was 37 (range 2-104) months. The 5-year OS rate of Lugano stagingⅠ, Ⅱ, Ⅲ and Ⅳ were 77.6%, 73.4%, 69.7%, 12.2% (χ(2)=63.395, P<0.001) respectively. The 5-year OS rate of TNM staging Ⅰ, Ⅱ, Ⅲ and Ⅳ were 77.6%, 75.9%, 25.0%, 9.3% (χ(2)=65.802, P<0.001) respectively. The 5-year OS rate of Musshoff stagingⅠ, Ⅱ, Ⅲ and Ⅳ were 84.5%, 68.4%, 25.0%, 9.3% (χ(2)=66.966, P<0.001) respectively. By Cox multiple-factors analysis, Lugano staging system was the only independent prognosis risk factor for PFS (HR=4.987, 95%CI: 1.421-17.498, P=0.009) and OS (HR=5.659, 95%CI: 1.563-20.485, P=0.008) of PGI-DLBCL. Univariated analysis revealed that the factors affecting PFS and OS of patients with PG-DLBCL include B-symptom, Eastern Cooperative Oncology Group performance status (ECOG PS), the number of extranodal lesions, serum lactate dehydrogenase (LDH), International prognostic index (IPI) score, staging and therapeutic regimen(all P values of PFS and OS<0.05). Patients with PG-DLBCL who received chemotherapy alone showed a better survival than others (PFS P=0.004; OS P<0.001); the factors affecting PFS and OS of patients with PI-DLBCL include ß2-microglobulin(ß2-MG), serum albumin(ALB) levels, LDH and staging (all P values of PFS and OS<0.05). Therapeutic regimen didn't affect those patients' survival (PFS P=0.661, OS P=0.720). The additional use of Rituximab failed to improve the survival of patients with PG-DLBCL and PI-DLBCL respectively (all P values of PFS and OS>0.05). Conclusions: Compared with TNM staging and Musshoff staging systems, Lugano staging system provides the best prognostic value in PFS and OS for patients with PGI-DLBCL. Accompany with B-sympto, higher ECOG PS score, more extranodal lesions, increased LDH, higher IPI score and later period are negative factors for PG-DLBCL. Increased ß2-MG and LDH, lower ALB level and later period are negative factors of PI-DLBCL.

[Block randomization in clinical trials].

Randomization is the key to ensure the balance of confounders between the comparison groups in clinical trials, and it is the statistical basis for making the study results comparable. A simple randomization in a clinical trial with large sample-size allows for a balanced comparison of the number of subjects and confounding factors between groups, but in a clinical trial with small sample-size, it is necessary to use a restricted randomization method (the blocked randomization). Block randomization ensures that the number of subjects between groups is basically equal, maximize the effectiveness of clinical trials as the standard error of the treatment-effect estimate is decreased, which affords big rewards in scientific accuracy and credibility.

[The ergonomic analysis of different holding gun methods for standing guard soldiers].

Objective: To study the effects of different holding gun methods and gun weight on health when standing guard, and propose a way to support the health of long-term standing guard soldiers. Methods: We created different percentile mannequins by Classic JACK, and adjusted the standing guard posture based on its standards for soldiers. The pressure on lumbar L4/L5 and moment on ankles and knees were analysied for different holding gun methods and gun weight. Then the mathematical models of joint load, gun weight and body mass index were studied by multiple regression analysis. Results: Holding gun methods and gun weight influence the force characteristics on ankles, knees and lumbar L4/L5. Holding gun with a brace and hands applying downward force -2 kgf could significantly reduce lumbar L4/L5 pressure. When the hand force is -5, -3, -4, -3, -2, -1, 0, 1, 2 kgf, and the weight of the gun is 0, Lumbar vertebrae L4/L5 joint pressure of people with different body mass index(P(1), P(5), P(55), P(95), P(99)) are the smallest, respectively 269, 281, 321, 408, 444 N, and the same change trend occurs when the weight of the gun is 2, 4, and 8 kg.The moment on ankles and knees were less with the same holding gun method and the hands downward force ranged from 0 to -4 kgf, and the higher the body mass index is, the more the hands downward force needed to make the moment on ankles and knees zero. That is, the moment on ankles could be zero when the hands downward force ranged from -1 to -3 kgf, the moment on knees could be zero when the hands downward force ranged from -1.1 to -3.7 kgf. Conclusion: To reduce the pressure on lumbar L4/L5 and moment on ankles and knees, so as to cut down occupational risk of long-standing operation, we advise the long-term standing guard soldiers holding gun with a brace and hands applying downward force -2 kgf.

[A clinic analysis of thoracolumbar vertebral fracture cascade].

Objective: To observe the clinical characteristics of thoracolumbar vertebral fracture cascade, analyze the relationship between the baseline fractures and the subsequent fractures and compare the distribution differences of subsequent fractures following vertebral augmentation or non-operation. Methods: From July 2012 to August 2016, 1 363 patients admitted to the First Affiliated Hospital of Soochow University with vertebral augmentation for the treatment of vertebral fractures were retrospectively analyzed.There were 190 cases of vertebral fracture cascade, 160 females and 30 males, with an average age of (74±9) years.The location and sequence of all vertebral fractures were recorded.The relationships between the baseline and the subsequent fractures were analyzed.According to different treatment on the baseline vertebral fractures, 190 cases were divided into vertebral augmentation group and non-operation group.The distribution differences of the subsequent fractures following vertebral augmentation and non-operation were compared with chi-square test. Results: Vertebral fracture cascade mainly located in the thoracolumbar spine T(11)-L(2) with an incidence of 52.0%.According to the direction of fracture development, the fracture cascade could be divided into up, down, centrifugation and concentration, and the incidence was 39.8%, 39.2%, 8.4% and 12.6%, respectively.The closer the vertebral body to the baseline fractures, the subsequent fractures incidence was higher.For distance with zero, one, two, three and four vertebrae, the incidence of subsequent vertebral fractures was 36.5%, 26.2%, 15.2%, 11.5% and 3.7%, respectively.A linear relationship was found between the subsequent fractures and the baseline fractures with a correlation coefficient of 0.90.The distribution difference of subsequent fractures between vertebral augmentation and non-operation group was not significant (χ(2)=17.16, P>0.05). Conclusions: The main directions of vertebral fracture cascade is up or down spiral development.The closer the vertebral body to the baseline fractures, the subsequent fractures incidence is higher.Vertebral augmentation doesn't affect the distribution of subsequent fractures.

[Analysis on influencing factors of unprotected sexual behavior among men who have sex with men based on structural equation model].

Objective: To analyze the influencing factors and mechanism for unprotected sexual behavior of men who have sex with men (MSM) in China. Methods: MSM who are more than 16 years old and have had homosexual sexual behaviors in recent years were recruited from five cities (Tianjin, Harbin, Nanjing, Chongqing and Xi'an) with active MSM population during 2013-2014 through network, site recruitment and peer recommendation using convenient sampling method. 3 519 subjects were included in this study. The sexual behavior related information of MSM in five cities was investigated using questionnaires. The confirmatory factor analysis was used to construct the measurement model by using the robust estimation method based on the weighted least square method to estimate parameters, and the final model was determined through evaluation and modification. Results: A total of 3 519 subjects were (29.9±8.76) years old, with a clear sexual orientation of 3 223 (91.6%), and 2 287 (65.0%) were aware of free HIV/AIDS treatment policies. The proportion of using condom last time was 77.2% (n=2 718), and the proportion of using condom every time was 36.9% (n=1 299) and 43.2% (n=1 521) respectively for the past six months with fixed and temporary partners. Structural equation model provided a good fit, the root mean square error of approximation, comparative fit index and non-normed fit index was 0.07, 0.97, 0.95, respectively. Knowing AIDS knowledge, educational level and sexual orientation had direct effects on unprotected sexual behavior, and the path coefficients were 0.179,-0.049 and -0.159, respectively. While the role of interventions was indirect, the path coefficient was 0.147. Conclusion: Lower education and unclear sexual orientation are disadvantages for reducing unprotected sexual behavior. Interventions mainly affect the unprotected sexual behavior by improving the level of HIV/AIDS knowledge. Increasing the comprehensive knowledge of MSM through intervention should be highly valued.

Treatment of Ribbing disease with 5-year follow-up and literature review.

Ribbing disease, or multiple diaphyseal sclerosis, is a rare diaphyseal sclerosis of unknown etiology. Patients with this pathology usually present with asymmetric pain limited to the lower extremities. Though all efforts are made to relieve the progressive pain associated with Ribbing disease, no medical or surgical treatments have been established yet. In this case report, we followed up a Ribbing case with sclerotic bone fenestration for 5 years. The radiological changes and the clinical effects are described, and the different Ribbing treatments are then briefly reviewed.

Retrospective analysis of 20 cases of refractory Hodgkin's lymphoma.

We investigate the clinical characteristics, prognosis and treatment of relapsed and refractory Hodgkin's lymphoma. Twenty patients with relapsed and refractory Hodgkin lymphoma were treated by chemotherapy or autologous stem cell transplantation in our hospital from April 2006 to August 2012. The retrospective analysis of the records from the 20 patients reflected both 5-year overall survival (OS) and progression-free survival (PFS). The overall effectiveness was 80% for the 20 patients. The 5-year overall survival rate and 5-year progression-free survival rate were 73.5% and 62.7%, respectively. Therefore, comprehensive treatment should be actively utilized in the case of invalid second-line regimen for the refractory HL patients.

The influence of testicular microlithiasis on the outcomes of in vitro fertilisation in a Chinese Han population.

To investigate the influence of testicular microlithiasis (TM) on the outcomes of in vitro fertilisation (IVF), we retrospectively reviewed the ultrasonography results of the TM patients who underwent IVF treatment in our hospital. They were classified as classic-TM (CTM) or limited-TM (LTM) comparing with patients without TM (Non-TM). Semen parameters, rates of fertilisation, normal fertilisation, good embryos and clinical pregnancy were analysed. The Non-TM group showed higher percentage of morphologically normal spermatozoa than the CTM or LTM group (4.08 ± 2.07 versus 3.40 ± 2.00 versus 3.04 ± 1.66, p = .003) while the other general semen parameters were comparable. The LTM group showed higher rate of fertilisation than the CTM group (85.10% versus 81.12%, p = .044). Moreover, the rates of normal fertilisation of Non-TM (62.47%) and LTM (66.32%) group were significantly higher than the CTM (58.02%) group (p = .027 and p = .001 respectively). There were 333 embryo transfer cycles in total (including 222 fresh and 111 frozen). The overall clinical pregnancy rate was 54.95%, 58.33% and 64.12% in the group of CTM, LTM and Non-TM respectively. However, no statistically significant difference was obtained (p = .326). Our results suggest that TM may have influence on the IVF outcomes. The extent of microlithiasis correlates inversely with the rates of fertilisation and normal fertilisation.

[Expression and prognostic significance of microenvironment related prognostic factors in patients with classical Hodgkin's lymphoma].

Objective: To observe the expression of three microenvironment related prognostic factors, i. e. programmed death 1 (PD-1), forkhead box protein 3(FOXP3) and colony-stimulating factor 1 receptor(CSF-1R) protein in classical Hodgkin's lymphoma (CHL) patients, and to explore the correlation between the protein expression and the prognosis of the patients. Methods: A total of 45 cases of CHL patients, who had been admitted to the Tianjin Medical University Cancer Institute and Hospital and Chinese PLA General Hospital from February 2005 to August 2010 were analyzed, including clinical features, prognostic factors, and treatment regimens. CHL patients' specimens were collected and the expression of PD-1, FOXP3, and CSF-1R proteins analyzed by immunohistochemical staining. Epstein-Barr virus encoded mRNA (EBER) was detected by in situ hybridization analysis. The relationship between the protein expression of PD-1, FOXP3 and CSF-1R and the patients' outcome was analyzed with clinical and follow-up data. Survival analysis was performed by Kaplan-Meier method, the Cox proportional hazard model was used to perform multivariate analysis. Results: In this cohort of 45 CHL patients, PD-1 positive was found in 7 cases (15.6%), FOXP3 high expression in 23 cases (51.1%), CSF-1R positive in 18 cases (40.0%). In the univariate analysis, the expression of FOXP3 and CSF-1R, International Prognostic Index (IPI) score, Ann Arbor stage and EBER were related with the patients' 5-year overall survival (OS); IPI score, the expression of FOXP3 and EBER were related with the patients' 5-year progress-free survival (PFS). Multivariate analysis indicated that CSF-1R protein expression was the independent prognostic factor affecting the patients' 5-year OS(HR: 8.918, P=0.020), and FOXP3 protein expression was the independent prognostic factor affecting the patients' 5-year PFS (HR: 0.122, P<0.001). And EBV was an independent prognostic factor of PFS and OS in the CHL patients. Conclusion: Microenvironment related prognostic factors FOXP3, CSF-1R and EBV may be independent prognostic factors of CHL and this study may provide novel strategies for targeted therapy of CHL.

[Clinical analysis of 9 cases with Anti-leucine-rich glioma inactivated 1 protein antibody associated limbic encephalitis].

Objective: This study was to describe the clinical characteristics of Anti-leucine-rich glioma inactivated 1 protein(LGI1) antibody associated limbic encephalitis. Methods: Clinical data including clinical features, laboratory and radiological findings, treatment and prognosis of the 9 patients were analyzed. Results: In all 9 cases, 6 cases experienced epileptic seizure, 5 cases had psychosis, 7 cases presented with memory impairment, 4 cases showed faciobrachial dystonic seizure, 2 had refractory hyponatremia. One case presented with typically acute Guillain-Barre syndrome (GBS). Anti-LGI1 antibody was detected in 6 cases in cerebrospinal fluid (CSF) samples and 9 in serum samples. Seven cases out of 9 had brain imaging abnormalities. All 9 cases found no evidence of tumors. Eight cases were given immune therapy. During a 1-16 months follow-up, 1 case had complete recovery, 5cases had various degree of sequelae , among whom 4 had memory disturbance and 1 case had changed personality. 2cases were lost to follow-up. Conclusions: Limbic encephalitis is the most common manifestation of anti-LGI1 antibody associated encephalitis. Faciobrachial dystonic seizure (FBDS) is a specific symptom which favors an early diagnosis. Tumor is uncommon to see. The disease has a relatively favorable prognosis while impaired memory can be seen as a common sequelae.

[Misdiagnosis and associated costs of benign paroxysmal positional vertigo].

Objective: The aims of this study were to investigate the misdiagnosis of benign paroxysmal positional vertigo (BPPV) and to estimate the associated costs. Methods: During October 2015 to December 2015, eighty patients were diagnosed with BPPV in the outpatient dizziness clinic of Shanghai Changzheng Hospital and the clinical data of all the 80 patients were collected including the demographic and clinical characteristics, the history of diagnosis, inappropriate diagnostic tests, costs of the medical tests, transportation and accommodation. All the data were investigated to estimate the misdiagnosis of benign paroxysmal positional vertigo and the associated costs in Shanghai, China. Results: This study showed that the misdiagnosis rate of BPPV was 60.0% (48/80) and the common inappropriate diagnostic tests for BPPV included Cranial CT and MRI test, cervical MRI, cervical and cerebrovascular investigations et al. There was no significant difference between the misdiagnosis patients (48) and patients without misdiagnosis (32) in gender, age, duration of symptom, involved canal and type of BPPV.Complications were significantly more frequent in the misdiagnosis group than for those without[81.3%(39 /48) vs 34.4%(11 /32)]. The estimated costs for each misdiagnosed individual were 8 502.98 China Yuan (CNY) and one-year economic burden associated with the misdiagnosis of BPPV in Shanghai was 13.184 7-78.862 1 million CNY. Conclusions: Our study suggests that the misdiagnosis rate of BPPV is high and the financial impact on patients and society with this disease is huge. The cost-effective Dix-Hallpike or supine roll test maneuver should be used before applying other expensive medical tests in order to minimize misdiagnosis and the waste of health care resources.

Circular RNA in blood corpuscles combined with plasma protein factor for early prediction of pre-eclampsia.

OBJECTIVE: To determine the expression of circular RNA (circRNA) in blood corpuscles of pregnant women before 20 weeks of pregnancy and to create a new model to identify the performance of circRNA combined with protein factors for the early diagnosis of pre-eclampsia (PE). DESIGN: Nested case-control study. SETTING: University medical centre, Guangzhou, China. POPULATION: A total of 1400 pregnant women recruited between 8 and 20 weeks of gestation. In all, 41 women with PE were included in the study, and were matched with 41 normally pregnant women based on maternal age and gestational age at same-size ratio. METHODS: The samples were analysed using a human circRNA microarray in the discovery phase, then the circRNA and the plasma protein factor endoglin (ENG) were validated. Finally we combined ENG with circRNA to create a new early prediction model for PE. MAIN OUTCOME MEASURES: Early changes of circRNA and ENG in PE. RESULTS: The circ_101222 levels in blood corpuscles of patients with PE were significantly higher than those in corresponding healthy women (P < 0.001). Using ENG in combination with circ_101222 resulted in a sensitivity of 0.7073, a specificity of 0.8049, and overall area under the curve of 0.876 (95% confidence interval 0.816-0.922) for the prediction of PE. CONCLUSION: CircRNA and plasma proteins may have some predictive value for PE (such as circ_101222 and ENG). The performance of each of these factors may be strengthened when plasma proteins are used in combination with circRNA. The results are preliminary and need to be validated in larger studies and other populations. TWEETABLE ABSTRACT: Plasma protein endoglin in combination with circ_101222 strengthened the predictive power for pre-eclampsia.

Engineering macrophages to control the inflammatory response and angiogenesis.

Macrophage (MΦ) dysregulation is increasingly becoming recognized as a risk factor for a number of inflammatory complications including atherosclerosis, cancer, and the host response elicited by biomedical devices. It is still unclear what roles the pro-inflammatory (M1) MΦ and pro-healing (M2) MΦ phenotypes play during the healing process. However, it has been shown that a local overabundance of M1 MΦs can potentially lead to a chronically inflamed state of the tissue; while a local over-exuberant M2 MΦ response can lead to tissue fibrosis and even promote tumorigenesis. These notions strengthen the argument that the tight temporal regulation of this phenotype balance is necessary to promote inflammatory resolution that leads to tissue homeostasis. In this study, we have engineered pro-inflammatory MΦs, MΦ-cTLR4 cells, which can be activated to a M1-like MΦ phenotype with a small molecule, the chemical inducer of dimerization (CID) drug. The MΦ-cTLR4 cells when activated with the CID drug, express increased levels of TNFα, IL-6, and iNOS. Activated MΦ-cTLR4 cells stay stimulated for at least 48h; once the CID drug is withdrawn, the MΦ-cTLR4 cells return to baseline state within 18h. Further, in vitro CID-activated MΦ-cTLR4 cells induce upregulation of VCAM-1 and ICAM-1 on endothelial cells (EC) in a TNFα-dependent manner. With the ability to specifically modulate the MФ-cTLR4 cells with the presence or absence of a small molecule, we now have the tool necessary to observe a primarily M1 MФ response during inflammation. By isolating this phase of the wound healing response, it may be possible to determine conditions for ideal healing.

Anti-apoptotic function of herpes simplex virus -2 latency-associated transcript RL1 sequence and screening of its encoded microRNAs.

BACKGROUND: The latency-associated transcript (LAT) gene of herpes simplex virus (HSV)-2 is the only detectable viral gene expressed during latent infection in neurons. LAT inhibits apoptosis and maintains latency by promoting the survival of infected neurons. However, whether LAT functions during HSV-2 infection via its encoded RNAs or via its encoded proteins remain unknown. Increasing evidence has indicated that LAT is likely to functionally promote the establishment of latent infection via LAT-encoded microRNAs (miRNAs). AIM: To explore whether the RL1 fragment of the five adjacent miRNAs has an effect on cell apoptosis, then provide supporting evidence to elucidate the potential role of these miRNAs and to aid screening of their cellular targets. METHODS: A number of techniques, including MTT assay, flow cytometry and DNA ladder analysis, were used to verify the role of the RL1 fragment and the contribution of the individual miRNAs to the anti-apoptotic effect. RESULTS: Five miRNAs (miR-H3, miR-H4-3p, miR-H4-5p, miR-H24 and miR-H19) were detected by quantitative PCR in PC12 cells stably expressing RL1 after pEGFP-RL1 plasmid transfection in vitro. The data indicated that expression of HSV-2 LAT RL1 seems to provide protection against apoptosis of PC12 cells induced by ActD. Antisense miRNAs specifically inhibiting these five miRNAs could efficiently reduce their expression. Transfection of antisense-miR-H3, antisense-miR-H4-5p and antisense-miR-H19 into PC12 cells stably expressing RL1 were able to partly reverse the anti-apoptotic effect of these miRNAs. CONCLUSIONS: These findings indicate that the apoptotic role of the RL1 fragment is likely to be related to overexpression of miR-H3, miR-H4-5p and miR-H19 in PC12 cells.

A novel method of real-time reverse-transcription loop-mediated isothermal amplification developed for rapid and quantitative detection of a new genotype (YHV-8) of yellow head virus.

UNLABELLED: A new genotype of yellow head virus (YHV), designated as YHV-8, was found in farmed shrimp Fenneropenaeus chinensis suffering suspectedly from EMS/AHPNS (early mortality disease/acute hepatopancreatic necrosis disease) in China in 2012. In this study, a one-step, real-time reverse-transcription loop-mediated isothermal amplification (rRT-LAMP) assay was developed for better detection of both genotypes of YHV-1 and YHV-8. A set of six specific primers was successfully designed targeting a conserved region of the YHV genome. The LAMP reaction was optimized to contain 8 mmol l(-1) Mg(2+) and 1·4 mmol l(-1) dNTPs, and to be performed at 58°C for 60 min. The detection sensitivity of the rRT-LAMP method was as low as 7 × 10(0)  copies per reaction. The specificity of the method was validated by the absence of any cross-reaction with the RNA samples extracted from other shrimp viruses (Taura syndrome virus, white spot syndrome virus, infectious hypodermal and haematopoietic necrosis virus, hepatopancreatic parvovirus) and specific pathogen-free (SPF) shrimp. The resulting standard curves showed high correlation coefficient values. Furthermore, the test of farm samples showed that YHV was detected in three of 111 Litopenaeus vannamei, six of eight Fenneropenaeus chinensis, five of 19 Macrobrachium rosenbergii and none of the nine Marsupenaeus japonicus. These results suggest that this assay is applicable widely as a new rapid and sensitive detection method in the research of YHV. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, we designate a new genotype of yellow head virus (YHV) as YHV genotype 8 (YHV-8) which was detected in diseased shrimp in China. A rapid, sensitive and specific rRT-LAMP detecting method for both YHV-8 and YHV-1 has been established. It is anticipated that this novel assay will be instrumental for diagnosis and surveillance of the virulent genotypes of YHV.