A multi-centre retrospective study of Nocardia speciation and antimicrobial susceptibility in Queensland, Australia.

The study aims to characterise the species identification and antimicrobial susceptibility testing (AST) results of Nocardial isolates from adult patients across major public hospitals in Queensland, Australia, over a 15-year period. A multi-centre retrospective observational study of Nocardia sp. isolates was conducted from 7 major public hospitals in Queensland, Australia, over a 15-year period. Clinical samples from patients aged ≥ 18 years that isolated Nocardia sp. were included. Demographic and clinical data were collected, along with species identification and AST results. Overall, 484 Nocardia sp. were isolated. Most patients were male (297, 61%) with a mean (IQR) age of 60 (51-75) and a median (IQR) Charlson Comorbidity Index of 4 (2-6). Of these, 239 (49%) patients were immunosuppressed. Organisms were most frequently isolated from sputum (174, 36%), and superficial swabs (102, 21%). Patients presented with pulmonary infections (165, 35%) and superficial skin and soft tissue infections (87, 18%) most commonly. One hundred (21%) isolates were deemed pulmonary colonisation and were not treated. Of the speciated organisms, N. nova complex was the most common (93, 19%), followed by N. farcinica complex (79, 16%). Organisms were reliably susceptible to linezolid (240/245, 98%), amikacin (455/470, 97%), and trimethoprim/sulfamethoxazole (459/476, 96%), but less so to imipenem (243/472, 51%) and ceftriaxone (261/448, 58%). This is the largest Australian description of Nocardia sp. to date. Given antimicrobials are often commenced prior to AST results and sometimes even speciation, characterisation of local species and antibiogram data is important to guide empiric choices and local guidelines.

Development and validation of a prediction model for unsuccessful treatment outcomes in patients with multi-drug resistance tuberculosis.

BACKGROUND: The World Health Organization has reported that the treatment success rate of multi-drug resistance tuberculosis is approximately 57% globally. Although new drugs such as bedaquiline and linezolid is likely improve the treatment outcome, there are other factors associated with unsuccessful treatment outcome. The factors associated with unsuccessful treatment outcomes have been widely examined, but only a few studies have developed prediction models. We aimed to develop and validate a simple clinical prediction model for unsuccessful treatment outcomes in patients with multi-drug resistance pulmonary tuberculosis (MDR-PTB). METHODS: This retrospective cohort study was performed between January 2017 and December 2019 at a special hospital in Xi'an, China. A total of 446 patients with MDR-PTB were included. Least absolute shrinkage and selection operator (LASSO) regression and multivariate logistic regression were used to select prognostic factors for unsuccessful treatment outcomes. A nomogram was built based on four prognostic factors. Internal validation and leave-one-out cross-validation was used to assess the model. RESULTS: Of the 446 patients with MDR-PTB, 32.9% (147/446) cases had unsuccessful treatment outcomes, and 67.1% had successful outcomes. After LASSO regression and multivariate logistic analyses, no health education, advanced age, being male, and larger extent lung involvement were identified as prognostic factors. These four prognostic factors were used to build the prediction nomograms. The area under the curve of the model was 0.757 (95%CI 0.711 to 0.804), and the concordance index (C-index) was 0.75. For the bootstrap sampling validation, the corrected C-index was 0.747. In the leave-one-out cross-validation, the C-index was 0.765. The slope of the calibration curve was 0.968, which was approximately 1.0. This indicated that the model was accurate in predicting unsuccessful treatment outcomes. CONCLUSIONS: We built a predictive model and established a nomogram for unsuccessful treatment outcomes of multi-drug resistance pulmonary tuberculosis based on baseline characteristics. This predictive model showed good performance and could be used as a tool by clinicians to predict who among their patients will have an unsuccessful treatment outcome.

P-incorporated CuO/Cu2S heteronanorods as efficient electrocatalysts for the glucose oxidation reaction toward highly sensitive and selective glucose sensing.

Currently, tremendous efforts have been made to explore efficient glucose oxidation electrocatalysts for enzymeless glucose sensors to meet the urgent demands for accurate and fast detection of glucose in the fields of health care and environmental monitoring. In this work, an advanced nanostructured material based on the well-aligned CuO/Cu2S heteronanorods incorporated with P atoms is successfully synthesized on a copper substrate. The as-synthesized material shows high catalytic behavior accompanied by outstanding electrical conductivity. This, combined with the unique morphology of unstacked nanorod arrays, which endow the entire material with a greater number of exposed active sites, make the proposed material act as a highly efficient electrocatalyst for the glucose oxidation reaction. Density functional theory calculations demonstrate that P doping endows P-doped CuO/Cu2S with excellent electrical conductivity and glucose adsorption capability, significantly improving its catalytic performance. As a result, a non-enzymatic glucose sensor fabricated based on our proposed material exhibits a broad linear detection range (0.02-8.2 mM) and a low detection limit (0.95 µM) with a high sensitivity of 2.68 mA mM-1 cm-2 and excellent selectivity.

[3D-printed vertebral body in anterior spinal reconstruction after total spondylectomy for patients with cervical chordoma].

OBJECTIVE: To investigate whether 3D-printed artificial vertebral body can reduce prosthesis subsidence rate for patients with cervical chordomas, through comparing the rates of prosthesis subsidence between 3D printing artificial vertebral body and titanium mesh for anterior spinal reconstruction after total spondylectomy. METHODS: This was a retrospective analysis of patients who underwent surgical treatment for cervical chordoma at our hospital from March 2005 to September 2019. There were nine patients in the group of 3D artificial vertebral body (3D group), and 15 patients in the group of titanium mesh cage (Mesh group). The patients' characteristics and treatment data were extracted from the medical records, including age, gender, CT hounsfield unit of cervical vertebra and surgical information, such as the surgical segments, time and blood loss of surgery, frequency and degree of prosthesis subsidence after surgery. Radiographic observations of prosthesis subsidence during the follow-up, including X-rays, CT, and magnetic resonance imaging were also collected. SPSS 22.0 was used to analysis the data. RESULTS: There was no significant difference between the two groups in gender, age, CT hounsfield unit, surgical segments, time of surgery, blood loss of posterior surgery and total blood loss. Blood loss of anterior surgery was 700 (300, 825) mL in 3D group and 1 500 (750, 2 800) mL in Mesh group (P < 0.05). The prosthesis subsidence during the follow-up, 3 months after surgery, there was significant difference between the two groups in mild prosthesis subsidence (P < 0.05). The vertebral height of the 3D group decreased less than 1 mm in eight cases (no prosthesis subsidence) and more than 1 mm in one case (mild prosthesis subsidence). The vertebral height of the Mesh group decreased less than 1 mm in five cases (no prosthesis subsidence), and more than 1 mm in eight cases (mild prosthesis subsidence). Two patients did not have X-rays in 3 months after surgery. There was a statistically significant difference between the two groups in the prosthesis subsidence rate at the end of 12 months (P < 0.01). The vertebral height of eight cases in the 3D group decreased less than 1 mm (no prosthesis subsidence) and one case more than 3 mm (severe prosthesis subsidence). Four of the 15 cases in the Mesh group decreased less than 1 mm (no prosthesis subsidence), two cases more than 1 mm (mild prosthesis subsidence), and nine cases more than 3 mm (severe prosthesis subsidence). There was a statistically significant difference between the two groups in the prosthesis subsidence rate at the end of 24 months (P < 0.01). The vertebral height of seven cases in the 3D group decreased less than 1 mm (no prosthesis subsidence), one case more than 3 mm (severe prosthesis subsidence), and one case died with tumor. One case in the Mesh group decreased less than 1 mm (no prosthesis subsidence), one case more than 1 mm (mild prosthesis subsidence), 11 case more than 3 mm (severe prosthesis subsidence), one case died with tumor and one lost the follow-up. Moreover, at the end of 12 months and 24 months, there was significant difference between the two groups in severe prosthesis subsidence rate (P < 0.01). CONCLUSION: 3D-printed artificial vertebral body for anterior spinal reconstruction after total spondylectomy for patients with cervical chordoma can provide reliable spinal stability, and reduce the incidence of prosthesis subsidence after 2-year follow-up.

Vision-Based Defect Inspection and Condition Assessment for Sewer Pipes: A Comprehensive Survey.

Due to the advantages of economics, safety, and efficiency, vision-based analysis techniques have recently gained conspicuous advancements, enabling them to be extensively applied for autonomous constructions. Although numerous studies regarding the defect inspection and condition assessment in underground sewer pipelines have presently emerged, we still lack a thorough and comprehensive survey of the latest developments. This survey presents a systematical taxonomy of diverse sewer inspection algorithms, which are sorted into three categories that include defect classification, defect detection, and defect segmentation. After reviewing the related sewer defect inspection studies for the past 22 years, the main research trends are organized and discussed in detail according to the proposed technical taxonomy. In addition, different datasets and the evaluation metrics used in the cited literature are described and explained. Furthermore, the performances of the state-of-the-art methods are reported from the aspects of processing accuracy and speed.

[Analysis the influence factors of treatment free remission outcome with chronic myeloid leukemia patients who discontinued tyrosine kinase inhibitors].

Objective: To explore the related factors affecting the outcome of treatment free remission (TFR) in patients with chronic myeloid leukemia (CML). Methods: Clinical data of CML patients with automatic discontinuation of tyrosine kinase inhibitor (TKI) from the CML cooperative organization of Henan province between June 2, 2013 to March 27, 2021 and the follow-up time was ≥ 6 months were retrospectively analyzed. Log-rank test was used for univariate analysis and Cox proportional risk regression model was used for multivariate analysis. Results: A total of 135 patients were enrolled, and 69 patients (51.1%) were femal and 66 patients (48.9%)were male. Median age was[M(Q1,Q3)] 49 years (38, 58)at discontinuation.Before discontinuation, 72 patients (53.3%) were on treatment with second-generation TKI, 63 patients (46.7%) were on treatment with IM, 17patients (12.6%) had a history of TKI reduction/withdrawal;median duration of treatment was months 84 (68, 108) for all patients;median time of TKI treatment to DMR was months 12(8, 26);median duration of DMR was months 65 (54, 84), and 9 patients (6.7%) had unsustained DMR.The median follow-up time was months 16(6-96), 35 patients (25.9%) lost MMR at a median months 3(1-22), overall estimated TFR was 74.1%.The univariate analysis results showed that:second-generation TKI was used, the time of TKI treatment to DMR was ≤12 months, DMR duration time ≥48 months, had sustained DMR, without TKI reduction/withdrawal history were favorable factors affecting of TFR in patients with TKI discontinuation (all P<0.05).The TFR rate of the second-generation TKI therapy group was significantly higher than the IM therapy group (81.9% vs 65.1%, P=0.019).The multivariate analysis results showed that second-generation TKI treatment[RR=0.451, 95%CI (0.227-0.896), P=0.023] and had sustained DMR [RR=0.120, 95%CI (0.053-0.271), P<0.001] were the protective factors of TFR in patients with TKI discontinuation. Conclusions: Treated with second-generation TKI and had sustained DMR are the protective factors of TFR in patients with TKI discontinuation.The CML patients who had sustained DMR more≥48 months before TKI discontinuation showed a better TFR.

Genome shuffling of Lactobacillus plantarum 163 enhanced antibacterial activity and usefulness in preserving orange juice.

Lactic acid bacteria have been used to inhibit the growth of spoilage bacteria in food and animal feeds. For instance, Lactobacillus plantarum 163 can inhibit efficiently the growth of both gram-positive and gram-negative bacteria. In our study, the antibacterial activity of L. plantarum 163 was further improved significantly by genome shuffling. The optimal conditions for protoplast formation and regeneration were 20 mg ml-1 lysozyme and 5 mg ml-1 mutanolysin for 30 min at 37°C using 0·5 mol l-1 sucrose as stabilizer. The protoplasts were inactivated under ultraviolet light for 120 s or heated at 58°C for 20 min. After two rounds of genome shuffling, the inhibitory activity of strain F2-14 was improved by 2·45- and 1·99-fold, respectively, as compared to their parent strains. The prepared antibacterial peptides supernatant (APS) was added to the orange juice to inhibit spores of Alicyclobacillus acidoterrestris (SAA) at 45 and 28°C. Results showed that the growth of A. acidoterrestris was significantly inhibited, and the decrease in total soluble solids, OD value and pH value was also delayed. After treatment with APS, the thermal sensitivity of spores was increased and its D value was reduced to 13·78, 3·87 and 1·47 min at 80, 90 and 95°C respectively.

Robust Korean License Plate Recognition Based on Deep Neural Networks.

With the rapid rise of private vehicles around the world, License Plate Recognition (LPR) plays a vital role in supporting the government to manage vehicles effectively. However, an introduction of new types of license plate (LP) or slight changes in the LP format can break previous LPR systems, as they fail to recognize the LP. Moreover, the LPR system is extremely sensitive to the conditions of the surrounding environment. Thus, this paper introduces a novel deep learning-based Korean LPR system that can effectively deal with existing challenges. The main contributions of this study include (1) a robust LPR system with the integration of three pre-processing techniques (defogging, low-light enhancement, and super-resolution) that can effectively recognize the LP under various conditions, (2) the establishment of two original Korean LPR approaches for different scenarios, including whole license plate recognition (W-LPR) and single-character license plate recognition (SC-LPR), and (3) the introduction of two Korean LPR datasets (synthetic data and real data) involving a new type of LP introduced by the Korean government. Through several experiments, the proposed LPR framework achieved the highest recognition accuracy of 98.94%.

[Acceptance evaluation of urine self-sampling, vaginal self-sampling and physician sampling in cervical cancer screening].

Objective: To compare the acceptance of urine self-sampling, vaginal self-sampling and physician sampling in cervical cancer screening. Methods: Questionnaire surveys were conducted in women attending cervical cancer screening in Xiangyuan County and Yangcheng County in Shanxi Province from July to November 2015 and from October to December 2018. Both surveys included the following three parts: feelings and acceptance of women in urine self-sampling, vaginal self-sampling and physician sampling, and the ease evaluation of two self-samplings. Results: There were 3 485 questionnaires were collected finally. The proportions of women felt embarrassed in urine self-sampling, vaginal self-sampling and physician sampling were 2.3% (81/3 472), 7.1% (247/3 472) and 8.6% (299/3 472), and 1.3% (46/3 472), 9.7%(3 37/3 472) and 14.9% (518/3 472) of women felt uncomfortable, and 1.0% (35/3 469), 8.5% (295/3 469) and 15.3% (531/3 469) felt pain, and 98.3% (3 334/3 393), 96.3% (3 267/3 393) and 99.0% (3 360/3 393) thought that the sampling were conducted properly (P<0.05). Based on the feeling during sample collection, 61.2% (1 876/3 064), 39.1% (1 199/3 064) and 66.5% (2 037/3 064) women were willing to use the corresponded sampling methods in cervical cancer screening, respectively (P<0.05). If the accuracy was the same, there were 31.8% (1 109/3 485) women preferred self-sampling and 68.2% (2 376/3 485) preferred physician sampling for cervical cancer screening. Meanwhile, 23.5% (820/3 482) preferred vaginal self-sampling in comparison with 76.5% (2 662/3 482) for urine self-sampling in cervical cancer screening. 86.5% (3 007/3 478) of the women thought urine self-sampling was very easy, comparing 40.9% (1 423/3 478) for vaginal self-sampling. Conclusions: The bad feeling of women during urine self-sampling is less common and less serious than those during physician sampling and vaginal self-sampling, and the acceptance for physician sampling is highest, following by urine self-sampling and vaginal self-sampling. Urine self-sampling is much easier than vaginal self-sampling.

Association between intraplaque neovascularization assessed by contrast-enhanced ultrasound and the risk of stroke.

AIM: To determine the cut-off value of the area ratio under the curve (ARUC) for predicting symptoms of stroke. MATERIALS AND METHODS: Contrast-enhanced ultrasound was used to analyse intraplaque neovascularization (IPN). The correlations between the ARUC and risk factors of stroke were examined. A receiver operating characteristic curve was used to determine the cut-off value of the ARUC. RESULTS: Using a quantitative analysis method for IPN, the ARUC was significantly higher in the symptomatic group than in the asymptomatic group (p=0.017). The ARUC was positively associated with the homocysteine level (r=0.429, p=0.002) and high-sensitivity C-reactive protein level (r=0.424, p=0.003). Regression analysis showed that the ARUC was a risk factor for symptoms of stroke. The receiver operating characteristic curve showed that the cut-off value for symptoms was 0.24; the sensitivity was 77%, and the specificity was 70%; the positive predictive value was 68%, and the negative predictive value was 78%. CONCLUSION: IPN was a risk factor for the occurrence of the clinical symptoms of stroke. Patients with an ARUC of >0.24 had a higher risk of stroke.

[The plasma level of amyloid-ß is associated with cognitive decline: a two years follow-up study in Xi'an rural areas].

Objective: To explore the relationship between plasma amyloid-ß (Aß) and cognitive decline during 2 year follow-up in a population-based cohort in Xi'an rural areas. Methods: The study was conducted in Qubao village in Xi'an suburbs cognitively normal residents over 40 years old were recruited from October 2014 to March 2015 and given a face-to-face standardized interview. Mini-mental state examination (MMSE) was employed to evaluate the global cognitive function, and quantification of plasma Aß was measured by sandwich enzyme-linked immunosorbent assay (ELISA) at baseline. Two years later, MMSE was tested at the end of study. Then logistic regression was performed to analyze the relationship between baseline Aß and cognitive change during 2 year follow-up. Results: A total of 1 020 participants completed the study, among whom 223 subjects (21.9%) presented MMSE scores decline (defined as MMSE scores decreased ≥2 points). Compared with those without decline, participants in the MMSE decline group were older (P<0.001) and had lower education level (P<0.001), while gender, hypertension, hyperlipemia, diabetes mellitus and APOE genotype were not significantly different between two groups. One-way analysis of variance (ANOVA) showed that the MMSE score decline was slighter in the lower tertile of baseline Aß(1)-40 compared with middle tertile (P=0.012), while MMSE decline were similar between different Aß(1)-42 level groups and Aß(1-42)/Aß(1-40) ratio groups (P=0.758, P=0.671, respectively). Multivariable logistic regression analysis showed that MMSE scores in the lower baseline plasma Aß(1-40) level declined more slowly (OR=0.565, 95%CI 0.379-0.845, P=0.005). However, the MMSE decline were also similar among different baseline plasma Aß(1-42) levels groups and Aß(1-42)/Aß(1-40) ratio groups. Conclusion: Population with lower level of baseline plasma Aß(1-40) manifests lower cognitive decline during 2 years, however further investigation on dynamics of plasma Aß and long term follow up are needed.

Combination of mRNA of Repair-related Genes in Rat Skeletal Muscles for Wound Age Estimation.

ABSTRACT: Objective To investigate the estimation of early and mid-term wound age by a combination of four mRNAs, the DNA polymerase delta-interacting protein 3 （POLDIP3） mRNA, regulator of chromosome condensation 1 like （RCC1L） mRNA, proline-rich 5 （PRR5） mRNA, and ribonucleic acid export 1 （RAE1） mRNA in rats skeletal muscles. Methods The model of rat skeletal muscle contusion was established, and then contusion area muscle tissue was extracted 4, 8, 12, 16, 20, 24, 28, 32, 36, 40, 44 and 48 h after injury. Histomorphological changes during the repair process after rat skeletal muscle contusion were observed. The relative expressions of Poldip3, Rcc1l, Prr5 and Rae1 mRNAs were detected by reverse transcription real-time quantitative polymerase chain reaction （RT-qPCR）. Different stages of wound age were classified by using the expression patterns of four genes at various time points after injury. The accuracy of the results was verified by Fisher discriminant analysis. Results Histomorphological results showed that the repair process after skeletal muscle contusion occurred with the prolonging of time. Through combination of the expression trends of the four kinds of mRNAs, the 48 h after injury could be divided into three periods, 4-12 h, 16-28 h and 32-48 h. The Fisher discrimination method showed that the classification accuracy rates of the three periods were 83.3%, 75.0% and 73.3%, respectively. Conclusion The classification discrimination based on the relative expression of every gene has a higher accuracy, and the accuracy of wound age estimation with combination of mRNA relative expressions is higher than that with a single indicator. By combining with Fisher discrimination method, this method can be used for early and mid-term wound age estimation.

[Real-world research on cervical cancer screening program and effect evaluation for Chinese population].

Objective: To evaluate the actual efficacy of cervical cancer and precancerous lesions screening approaches in real-world regions with different economic levels in China. Methods: The demonstrative application and effect evaluation of cervical cancer screening program were conducted in 21 hospitals nationwide from 2015 to 2018. Multi-stage sampling method was adopted to divide the country into 7 large areas according to geographical location. Two to four screening sites of two types of cancer (cervical cancer and breast cancer) were selected in each area, and the grassroots screening sites were implemented under the guidance of superior hospitals. In rural areas, women were initially screened using cytology, human papillomavirus (HPV) testing and visual inspection. The women with positive cytology or visual inspection were referred for colposcopy, and the women with positive HPV infection were randomly referred for reflex cytology or visual inspection, or direct colposcopy examination. In urban areas, women were primarily randomized into cytology or HPV testing groups. The women with abnormal cytology or positive HPV 16/18 infection were directly referred for colposcopy examination, whereas the women with positive infection of the other 12 high-risk subtypes of HPV were referred for reflex cytology or colposcopy. All of recruited women would be follow-up and screened by the baseline screening techniques in the third year while the positive women underwent colposcopy examination. The positive rates, referral rates, the detection rates of grade 2 and above of cervical intraepithelial neoplasia (CIN 2+ ) were compared. Results: A total of 63 931 women were recruited at the baseline. Among them, 11 rural sites included 33 823 women: 15 577, 11 157 and 7 089 women were screened by HPV testing, visual inspection via acetic acid or Lugol's iodine (VIA/VILI) and cytology, respectively. Additionally, 30 108 women were from 10 urban sites: 9 907 and 20 201 women were screened by cytology and HPV subtyping, respectively. The HPV positive rate for urban women was 9.34%, whereas that for rural women was 12.53%. The abnormal rate of cytology for urban women was 5.63%, and that for rural women was 4.24%. The positive rate of VIA/VILI in the rural women was 12.25% Furthermore, the detection rate of CIN2+ at the baseline was 0.56%, and that was statistically higher in HPV-positive group than cytology-positive group (P<0.05). Conclusions: All of screening sites completed the baseline screening tasks as expected. The prevalence of HPV infection is higher in rural women than urban women. The performance of HPV testing is better than cytology for detecting CIN2+ cases. This real-world demonstration study provides evidences for cervical cancer prevention and control in different regions.

[Treatment of subcutaneous fistula secondary to cerebrospinal fluid leakage in thoracic spinal stenosis cases].

OBJECTIVE: To investigate the treatment strategy for subcutaneous fistula secondary to cerebrospinal fluid leakage (CSFL) in thoracic spinal stenosis (TSS) cases. METHODS: In the study, 186 CSFL cases diagnosed with TSS and operated in general spine group of Department of Orthopedics, Peking University Third Hospital from January 2005 to December 2014 were retrospectively reviewed, of which eleven had subcutaneous fistula secondary to CSFL and were regularly followed up. Treatment strategy for subcutaneous fistula depended on the severity of CSFL and the recovery rate of thoracic myelopathy. Japanese Orthopedic Association (JOA) score was utilized to evaluate the neurologic status of these patients preoperatively and postoperatively. Statistical analysis was conducted between preoperative and postoperative JOA scores. RESULTS: All of the 11 patients were regularly followed up for at least 24 months. Six of them had ossification of the posterior longitudinal ligament (OPLL) combined with ossification of ligamentum flavum (OLF), all of them undertook "cave-in" 360° circumferential decompression of the spinal cord with instrumentation. Five cases had OLF only, and received En bloc resection of lamina and OLF and fixation. The follow-up period ranged from 30 months to 131 months, and averaged at (85±34) months. Preoperative symptoms lasted from 3 months to 8 years, and the median was 18 months. Drainages were placed for 2-6 days, and averaged at (4.2±1.1) days. Ten cases appeared with fever during the perioperative period, the maximum body temperature was (37.3-39.7) °C. Prolonged antibiotics were applied in two cases with high fever. Ten cases were treated with conservative methods, CSFL were completely absorbed during the follow-up time, of which compressive dressing was utilized in 8 cases, and punctures combined with compressive dressing were used in 2 cases. For only 1 case, conservative therapy failed and reoperation was required because of neurological deterioration arising from CSF pseudocyst. For these 11 cases, preoperative JOA score arose from (3.8±1.6) preoperatively to (8.9±1.2) at the end of the final follow-up, the recovery rate was 70.8%. No infection of wound or central nerve system were noticed, and neither were unhealing wound. CONCLUSION: Most TSS cases with subcutaneous fistula secondary to CSFL could be cured by conservative methods, and reoperation is required only if myelopathy caused by cerebrospinal fluid pseudocyst is identified.

[Study on effect and mechanism of Honokiol enhancing tumor necrosis factor-related apoptosis inducing ligand against hepatocellular carcinoma HepG2 cells via activating JNK signaling pathway].

Objective: To investigate the effect and intrinsic mechanism of Honokiol (HNK) enhanced tumor necrosis factor-related apoptosis inducing ligand (TRAIL) on hepatocellular carcinoma HepG2 cells. Methods: HepG2 cells were routinely cultured. Firstly, a concentration gradient of HNK was given to observe its effect on the vitality of tumor cells. Western blot were used to detect change in the expression levels of c-jun N-terminal kinase (JNK), death receptor 4 (DR4), and DR5.Secondly, we observed the effect of combined drugs (HNK and TRAIL) on the vitality of tumor cells. Apoptosis-related protein expression levels were detected to determine the apoptosis condition. Thirdly, JNK inhibitor SP600125 was used to block the JNK pathway, and it was evaluated whether JNK signaling pathway mediated the DR4 and DR5 levels and finally, the subcutaneous tumor model of nude mice was constructed, and enhancement effect of HNK on TRAIL was confirmed in vivo. Results: Cell vitality was decreased (P < 0.05) in a dose-dependent manner after treatment with gradient HNK. Combined effect of TRAIL and HNK was superior to that of single drug administration (P < 0.05). Western blot analysis showed that pJNK level increased after HNK treatment and TRAIL receptor DR4 and DR5 expression were up-regulated. Combined application of HNK and TRAIL, B lymphocyte tumor factor 2 (BCL2) decreased significantly while Bcl2 related X protein (Bax) increased significantly. Blocking JNK pathway by SP600125, the expression of DR4 and DR5 decreased (P < 0.05), Bax expression decreased and Bcl2 expression increased compared with HNK+TRAIL group. In vivo results showed that TRAIL inhibited the growth of subcutaneous tumors, and enhanced inhibition effect in combination with TRAIL and HNK. Conclusion: HNK may enhance the inhibitory effect of TRAIL on HepG2 cells by activating JNK pathway and then upregulating the expression of DR4 and DR5.

[Effects of reconstruction with unicondylar osteoarticular allografts with or without prosthesis for bone tumors around knee joint].

Objective: To investigate the survival rate, function outcomes, and complications after using unicondylar osteoarticular allografts with or without prosthesis to reconstruct the knee joint for tumors located in distal femoral or proximal tibial uni-condyle. Methods: Twenty-two patients who underwent unicondylar osteoarticular allografts with or without prosthesis composite reconstructions from January 2007 to December 2015 in Department of Orthopaedic Surgery of Xi Jing Hospital, the Fourth Military Medical University were retrospectively reviewed. There were 14 males and 8 females and the mean age was 35 years(8-65 years). There were 12 malignent tumors and 10 aggressive benign tumors. The tumors were located in distal femur in 14 cases and proximal tibia in 8 cases. After tumor excision, the distal femur was reconstructed with unicondylar osteoallograft-prosthesis composite, and proximal tibial plate was reconstructed with unicondylar osteoarticular allograft with the help of computer-assisted navigation system. Function and radiograph were documented according to the Musculoskeletal Tumor Society (MSTS) functional scoring system and the International Society of Limb Salvage (ISOLS) radiographic scoring system. The median follow-up time was 60 months (5-116 months). Results: At the latest follow-up, 2 patients had amputation owing to local recurrence in 12 malignant tumors. Three patients had pulmonary metastasis and 1 patient died another 2 alive with disease. Kaplan-Meier analysis indicated that the disease-free survival rate was 73%. There was no recurrence and metastasis in 10 patients with giant cell tumor. The average MSTS score was 26 points and the radiographic score was 78%-94%(average 90%). The complications included superficial infection in 1 patient and screw broken in 1 patient. There was no broken or collapse allograft in all composite reconstruction patients but 6 cases in allograft reconstruction. Conclusions: Unicondylar osteoarticular allografts or prosthesis composite was feasible reconstruction for tumors in distal femoral uni-condyle. It could provide good functional outcomes and also prevent joint degeneration. Similarly, allograft reconstruction was also a reliable technique for proximal tibial defect.

IDH mutations in cancer and progress toward development of targeted therapeutics.

Isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) are key metabolic enzymes, converting isocitrate to α-ketoglutarate (αKG).IDH1 and IDH2 mutations have been identified in multiple tumor types, including gliomas and myeloid malignancies such as acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). Here we provide an overview of the function of normal and mutated IDH, discuss the role of IDH mutations in tumorigenesis and progression and review the key clinical considerations when treating IDH-mutated tumors based on emerging clinical data from mutant IDH1/2 inhibitor trials. IDH1 and IDH2 mutations confer neomorphic activity in the mutant protein, resulting in the conversion of αKG to the oncometabolite, D-2-hydroxyglutarate (2-HG). The subsequent accumulation of 2-HG results in epigenetic dysregulation via inhibition of αKG-dependent histone and DNA demethylases, and a block in cellular differentiation. There is growing preclinical and clinical evidence suggesting that IDHmutations are involved in neoplasia. Furthermore, preclinical studies assessing small molecule inhibitors of mutant IDH1/2 enzymes have provided proof of concept that this approach decreases intracellular 2-HG levels, reverses epigenetic dysregulation and induces cellular differentiation. Phase I studies of mutant IDH inhibitors are currently ongoing in patients with IDH-mutant hematologic and solid tumors, with early data in hematologic tumors suggesting a manageable safety profile as well as clinical benefit, with a mechanism of action based on differentiation of malignant cells. Inhibition of mutant IDH shows promise as a treatment approach in hematologic malignancies, with further development ongoing in solid tumors and glioma. The mutant IDH inhibitors may have clinical utility both as single agents and in combination strategies that target additional oncogenic pathways.

MiR-21 functions oppositely in proliferation and differentiation of neural stem/precursor cells via regulating AKT and GSK-3ß.

MicroRNA involves in regulating behavior of neural stem/precursor cells (NSPCs), thus it offers the potential to treat central nervous system disease. However, the effect of miR-21 on NSPCs remains unknown. In this study, we demonstrated that miR-21 reduced proliferation and promoted neural differentiation in NSPCs via regulating the activation of AKT and GSK-3ß signaling pathways in vitro. During differentiation of NSPCs, the expression of miR-21 was increased in a time-dependent manner by qRT-PCR. Synthesized pre-miR-21 or anti-miR-21 was transfected into NSPCs, thereby efficiently overexpressing or knocking down miR-21. Overexpression of miR-21 promoted the neural differentiation of NSPCs, as indicated by Tuj1 and PSA-NCAM staining. Interestingly, knocking down miR-21 had the opposite effect of neural differentiation in NSPCs. However, in proliferation area, overexpression of miR-21 decreased the cell viability by 3-((2-methyl-1,3-thiazol-4-yl)ethynyl)pyridine hydrochloride (MTT) assay, and inhibited the proliferation of NSPCs, as indicated by 5-Bromo-2-deoxyUridine (BrdU) staining. And likewise, knocking down miR-21 had the opposite effect of cell viability and proliferation. Western blot showed that overexpression of miR-21 enhanced the expression of Cyclin D1, however, knocking down miR-21 prevented its expression. Furthermore, we revealed that protein kinase B (AKT) and glycogen synthase kinase-3 beta (GSK-3ß) signaling pathways were involved in the proliferation and neural differentiation of NSPCs. Overexpression of miR-21 activated AKT, and the p-GSK-3ß was increased. Conversely, knocking down miR-21 blocked the activation of AKT, and decreased the phosphorylation level of GSK-3ß. These results demonstrated that miR-21 promotes neural differentiation and reduces proliferation in NSPCs via regulating AKT and GSK-3ß pathways. These findings may help to develop strategies for treatment of central nervous system diseases.

Population structure analysis and association mapping of blast resistance in indica rice (Oryza sativa L.) landraces.

Rice blast caused by Magnaporthe oryzae is one of the most devastating rice diseases worldwide. To understand the genetic diversity of indica landrace accessions and identify simple sequence repeat (SSR) markers that are associated with blast resistance, a population of 276 indica landraces from across the world was constructed. This population was then used to evaluate the blast-resistance phenotype through artificial inoculation under controlled conditions in 2012 and 2013. The genetic diversity and association of the population with resistance were analyzed by examining the phenotype for 160 SSR markers distributed on 12 rice chromosomes. The 276 accessions were classified into seven groups using model- and distance-based cluster analyses. Associations between SSR markers and blast resistance showed that 26 SSR markers were significantly associated with blast resistance in 2012 and 2013 (P < 0.01) and that the phenotypic variation ranged from 2.68 to 13.11%. Nineteen of the markers associated with blast resistance were located in regions where genes or quantitative trait loci (QTLs) have been previously reported, and seven were newly identified in this study. These results indicate that marker-trait association has potential advantages over classical linkage analysis and QTL mapping, and that these markers could be used for marker-assisted selection in rice blast-resistance-breeding programs.

Appropriate Osmotic Balance Duration for Different Volumes of Ovarian Tissue in Vitrification Solution: a Study of Ovary Tissue Vitrification and Transplantation in Sheep.

BACKGROUND: Auto-transplantation of cryopreserved ovarian tissue has become a promising method for fertility preservation and standardization of the process is crucial for practical applications. OBJECTIVE: Here we used different size of large sheep ovarian cortex to study the most suitable osmotic balance durations in the vitrification solution for large piece ovary cortex. MATERIALS AND METHODS: The ovarian cortices from six-month old female sheep were divided into 40 cubic mm, 80 cubic mm and 160 cubic mm volume, A two-step osmotic balance method was used based on the results from morphological and histological study, we detected the expression of VEGF after thawing, the percentage of follicles that survived and serum E2 levels,together with apoptosis test by TUNEL. RESULTS: the morphology of follicles and stromal cells were the best when the osmotic balance duration was 7 min, 11 min and 19 min, respectively. Osmotic balance time(T) is proportional to the area(S), we deduced that areas(S) of large pieces of ovarian cortex in vitrification fluid conformed to the standardized osmotic balance time(T) formula; i.e., T=（S+15)/5 (22~24C, sample thickness 1-2 mm), In vitro culture of vitrification-thawed large cortex ovaries and xenogenic heterotopic transplantation by using the standardized osmotic balance duration formula were both successful. CONCLUSION: Formula T=（S+15)/5 can be used to calculate optimal osmotic balance duration for different size of ovarian cortexes during vitrification.