[The predictive value of dynamic changes of neutrophil/lymphocyte ratio (NLR) combined with the model of end-stage liver disease (MELD) score in patients with acute-on-chronic hepatitis B liver failure].

Objective: To determine the predictive value of dynamic changes of neutrophil/lymphocyte ratio (NLR) combined with the model of end-stage liver disease (MELD) score in patients with acute-on-chronic hepatitis B liver failure. Methods: Patients with acute-on-chronic hepatitis B liver failure who were hospitalized in the Department of Hepatology of Qilu Hospital of Shandong University from January 2010 to July 2023 were retrospectively enrolled. According to the clinical outcomes of patients within 30 days of admission, they were divided into the survival group and the death group. The dynamic changes in NLR and initial values on day 3, 5, 8, and 12 in two groups were analyzed for the diagnostic value of 30-day prognosis in patients with acute-on-chronic hepatitis B liver failure. Logistic regression analysis and machine learning XGBoost algorithm were used to evaluate the risk factors influencing the prognosis of patients at 30 days. Receiver operating characteristic(ROC) curve was used to evaluate the diagnostic value of NLR and initial value change combined with MELD score on day 12 of admission in patients with chronic acute hepatitis B liver failure. Results: A total of 243 patients were enrolled in the study, including 145 patients in the survival group [115 males, 30 females, aged 25-74 (47±11)] and 98 patients in the death group [80 males, 18 females, aged 22-80 (49±13) ]. The median initial NLR of survival group and death group were 3.5 (2.1, 5.3) and 4.9 (2.9, 8.3), respectively, and the difference was statistically significant (P=0.003). The variation of NLR from the initial value on day 3, 5, 8, and 12 in the survival group [1.6 (0, 4.3), 1.9 (-0.2, 4.1), 2.0 (-0.1, 4.3) and 2.9 (0.3, 7.0), respectively] were lower than that in the death group [3.2 (0.9, 7.5), 5.1 (1.8, 7.6), 5.8 (2.0, 10.6) and 9.6 (3.5, 16.4), respectively] (all P<0.001). Logistic regression multivariate analysis showed that the changes in NLR on the 12th day and initial value (OR=1.07,95%CI:1.01-1.14, P=0.014), the changes in NLR on the 3rd day and initial value (OR=2.71, 95%CI: 1.32-5.55, P=0.007), the initial value of NLR (OR=1.18,95%CI:1.01-1.37,P=0.035) and fibrinogen (OR=0.21,95%CI:0.05-0.96,P=0.044) were related factors for death within 30 days. Machine learning XGBoost algorithm showed that the weight of the change between the NLR on the 12th day and the initial value was the highest. The area under the ROC curve of the combined MELD score was 0.812 (95%CI: 0.728-0.895), the specificity was 67.78%, and the sensitivity was 82.35%. Conclusion: Dynamic change of NLR combined with MELD score has high predictive value for the short-term prognosis of patients with acute-on-chronic hepatitis B liver failure.

Liquid-like atoms in dense-packed solid glasses.

Revealing the microscopic structural and dynamic pictures of glasses is a long-standing challenge for scientists1,2. Extensive studies on the structure and relaxation dynamics of glasses have constructed the current classical picture3-5: glasses consist of some 'soft zones' of loosely bound atoms embedded in a tightly bound atomic matrix. Recent experiments have found an additional fast process in the relaxation spectra6-9, but the underlying physics of this process remains unclear. Here, combining extensive dynamic experiments and computer simulations, we reveal that this fast relaxation is associated with string-like diffusion of liquid-like atoms, which are inherited from the high-temperature liquids. Even at room temperature, some atoms in dense-packed metallic glasses can diffuse just as easily as they would in liquid states, with an experimentally determined viscosity as low as 107 Pa·s. This finding extends our current microscopic picture of glass solids and might help establish the dynamics-property relationship of glasses4.

[Analysis of common viral infection in surveillance cases of febrile respiratory syndrome in 9 provinces of China from 2009 to 2021].

Objective: To understand the common viral infection among the surveillance cases of fever respiratory syndrome (FRS) in nine provinces in China. Methods: The research data were obtained from nine provinces (Anhui, Beijing, Guangdong, Hebei, Hunan, Jilin, Shandong, Shaanxi and Xinjiang) in the "Infectious Disease Surveillance Technology Platform Information Management System" of the Chinese Center for Disease Control and Prevention from January 2009 to June 2021. Finally, 8 243 FRS cases with nucleic acid detection results of eight viruses [human influenza virus (HIFV), human respiratory syncytial virus (HRSV), human adenovirus (HAdV), human parainfluenza virus (HPIV), human rhinovirus (HRV), human metapneumovirus (HMPV), human coronavirus (HCoV) and human Boca virus (HBoV)] were included in the study. The χ2 test/Fisher exact probability method was used to analyze the difference of virus detection rate in different age groups, regions and seasons. Results The M (Q1, Q3) age of 8 243 FRS cases was 4 (1, 18) years old, and 56.56% (4 662 cases) were children under 5 years old. Males accounted for 58.1% (4 792 cases) of all cases. All cases were from outpatient/emergency department (2 043 cases) and inpatient department (6 200 cases). The virus detection rates of FRS cases from high to low were HRSV, HIFV, HPIV, HRV, HAdV, HMPV, HCoV and HBoV. Two or more viruses were detected simultaneously in 524 cases, accounting for 15.66% of virus-positive cases. The difference of the virus detection rate in different age groups was statistically significant (all P values<0.05), and the virus detection rate in children<5 years old was higher (49.96%). The positive rate of any virus in south China was higher than that in north China (P<0.001). The virus-positive FRS cases were detected throughout the year. The detection rate of HRSV was higher in autumn and winter. The detection rate of HIFV was higher in winter. The detection rate of HMPV was higher in winter and spring. The detection rates of HPIV, HRV, HCoV and HBoV were higher in summer and autumn, while there was no significant difference in the detection rate of HAdV in different seasons. Compared with 2009-2019, the detection rate of any virus in 2020-2021 decreased from 41.37% to 37.86%. The detection rate of HIFV decreased sharply from 10.62% to 1.37%. The detection rate of HPIV decreased from 8.24% to 5.88%. The detection rate of HRV and HBoV increased from 5.43% and 1.79% to 9.67% and 3.19%, respectively. Conclusion: HRSV and HIFV infections are more common among FRS cases in nine provinces in China from 2009 to 2021, and the epidemiological characteristics of eight common respiratory viruses vary in different age groups, regions and seasons.

[Role of crenel lateral lumbar interbody fusion in the precise selection of fusion level in posterior correction for adult degenerative scoliosis].

Objective: To investigate the role of minimally invasive crenel lateral lumbar interbody fusion (CLIF) in the decision of fusion level in posterior correction for severe adult degenerative scoliosis. Methods: This is a prospective study.Patients with level Ⅴ and Ⅵ of Lenke-Silva classification who were treated at Department of Orthopedics,the Second Affiliated Hospital, School of Medicine, Zhejiang University from June 2016 to March 2019 were included.First,the enrolled patients completed the preoperative clinical and imaging examination,the Lenke-Silva classification was evaluated,the surgical segments in first-stage CLIF was determined and the fusion segments required for single-stage posterior correction was predicted.After the first-stage CLIF,patients received reassessment of Lenke-Silva classification and global coronal and sagittal balance.Patients were divided into two groups:the effective group (level of Lenke-Silva classification decreased) and the ineffective group (level of Lenke-Silva classification unchanged).Second-stage posterior surgery was performed based on the results of reassessments.The fusion segment,Cobb angle,parameters of global coronal and sagittal plane,visual analogue pain score (VAS) and Oswestry disability index (ODI) were compared between the two groups preoperatively,after first-stage CLIF,second-stage posterior fixation and at the final follow-up.The potential factors associated with the decrease of the level of Lenke-Silva classification were recorded and compared between the two groups.Independent sample t test,repeated measure analysis of variance,rank sum test,χ2 test or Fisher exact method were used to compare the difference among groups. Results: Fifty-four patients were enrolled,including 8 males and 46 females,aged (68.8±5.8) years (range:56 to 77 years).Preoperatively,26 patients were classified as level Ⅴ by Lenke-Silva classification,28 cases were grade Ⅵ.CLIF was performed in 194 segments,with 114 segments(58.8%) receiving anterior column realignment (ACR) and 15 segments(7.7%) using hyperlordotic cages.After first-stage CLIF,22 patients with level Ⅴ and 10 patients with Ⅵ of Lenke-Silva classification decreased and were classified into effective group.The level of the remaining 4 patients with level Ⅴ and 18 patients with grade Ⅵ unchanged and were classified into ineffective group.Preoperatively,the apical vertebrae was below L1 in all 32 patients of effective group and 18 (81.8%,18/22) patients of ineffective group.The difference was statistically significant (P=0.023).There were 7 (31.8%,7/22) patients had continuous osteophyte in front of the intervertebral space in ineffective group,while none patient had continuous osteophyte in front of the intervertebral space in effective group,and the difference was statistically significant (P=0.001).In first-stage CLIF,more intraoperative ACR(71.2% vs.39.5%,χ²=20.660,P<0.01)and hyperlordotic cage (12.7% vs.0,P=0.001) were used in the effective group,while there was less severe cage subsidence after the operation (5.9% vs.15.8%,χ²=4.793,P=0.029) in effective group.After first-stage CLIF,there was no difference in the Cobb angle between the two groups.While,lumbar lordosis (LL) in effective group (34.0±8.3)° was greater than that of the ineffective group (25.5±9.7)° (t=3.478,P=0.001),and the difference between the pelvic incidence (PI) and LL in effective group (15.7±4.6)°was significantly smaller than ineffective group(20.0±10.8)° (t=-2.129,P=0.038).The posterior fusion levels was less,the rate of fusion to thoracic spine region and the actual fusion segment was less than that of single-stage posterior correction in effective group (all P<0.01).All patients were follow-up for 24 to 45 months.There was no significant difference in radiological and clinical results between the two groups after first-,second-stage surgery and at the final follow-up (all P>0.05). Conclusions: First-stage CLIF decreased the Lenke-Silva classification of some patients with severe degenerative scoliosis.Combined with the reassessment of Lenke-Silva classification and global coronal and sagittal plane,it helps to accurately determine the fusion segment.Decrease of Lenke-Silva classification is associated with the preoperative level of apical vertebrae,continuous osteophytes in front of the intervertebral space,intraoperative use of ACR and hyperlordotic cage and the degree of cage subsidence postoperatively.

[Analysis of families with fetal congenital abnormalities but negative prenatal diagnosis by whole exome sequencing].

Objective: To evaluate the value of whole exome sequencing (WES) in prenatal clinical application. Methods: A total of 1 152 cases of congenital abnormal [including structural malformation, nuchal translucency (NT) thickening and intrauterine growth restriction] with traditional prenatal diagnosis [including G-band karyotype analysis and chromosome microarray analysis (CMA)] negative were analyzed. The congenital abnormal fetuses were divided into retrospective group and prospective group according to the time of WES detection, that is whether the pregnancy termination or not. According to the specific location of fetal malformation and their family history, the cohort was divided into subgroups. The clinical prognosis of all fetuses were followed up, and the effect of WES test results on pregnancy decision-making and clinical intervention were analyzed. According to the follow-up results, the data of fetuses with new phenotypes in the third trimester or after birth were re-analyzed. Results: Among 1 152 families who received WES, 5 families were excluded because of nonbiological parents. Among the remaining 1 147 families, 152 fetuses obtained positive diagnosis (13.3%,152/1 147), including 74 fetuses in the retrospective group (16.1%,74/460) and 78 fetuses in the prospective group (11.4%,78/687). In fetuses with negative CMA and G-band karyotype analysis results but new phenotypes in the third trimester or after birth, the positive rate by WES data re-analysis was 4.9% (8/163). A total of 34 (21.3%, 34/160) fetuses were directly affected by the corresponding positive molecular diagnosis. Among 68 cases of live births with diagnostic variation grade 4, 29 cases (42.7%, 29/68) received appropriate medical intervention through rapid review of WES results. Conclusions: WES could increase the detection rate of abnormal fetuses with negative G-banding karyotype analysis and CMA by 13.3%. Prenatal WES could guide pregnancy decision-making and early clinical intervention. It might be an effective strategy to pay attention to the special follow-up of the third trimester and postnatal fetus and to re-analyze the WES data.

Controlled synthesis of nanostructured glassy and crystalline high entropy alloy films.

High entropy alloy (HEA) based thin films have been attracting increasing research interest recently because of their unique mechanical/physical properties. However, the physical mechanisms that govern the formation of the atomic structure in HEA thin films are not clear yet. In this work, we synthesized a series of FeCoNiNb0.5 HEA thin films via direct current magnetron sputtering with carefully controlled processing parameters. Through a systematical study by x-ray diffraction and transmission electron microscopy, we demonstrated that the atomic structure of the HEA thin films of the same composition could exhibit different nanostructures and metastable phases, including amorphous and metastable crystalline phases. In addition, we also developed a physical model which sheds quantitative insights into the thermodynamics and kinetics for the phase selection in our HEA thin films. Our current work could pave a way for a controlled synthesis of a variety of nanostructured chemically complex alloy thin films for future structural and functional applications.

Efficacy of bioactive nanoparticles on tissue-endotoxin induced suppression of stem cell viability, migration and differentiation.

AIM: To characterize a lipopolysaccharide (LPS)-treated dentine tissue model (LPS dentine) to analyse the efficacy of polycationic chitosan nanoparticles (CSnp) and/or dexamethasone conjugate chitosan nanoparticles (Dex-CSnp) on the viability/differentiation potential of stem cells from apical papilla (SCAP) when exposed to LPS dentine. A further aim was to understand the effect of macrophage-dependent inflammation on SCAP migration in the presence of LPS dentine. METHODOLOGY: A total of 88 dentine slabs were used. TOF-SIMS analysis was performed amongst the LPS-treated and untreated dentine groups (n = 2/group). The study was conducted using four dentine groups: no treatment (control); LPS treatment only; LPS treatment followed by CSnp conditioning; and LPS treatment followed by Dex-CSnp conditioning groups. SCAP adherence, viability, differentiation and biomineralization potential on dentine from different groups were studied using fluorescent and scanning electron microscopy. Inflammation by macrophages in response to LPS dentine was quantified, and effect on SCAP migration was analysed. Statistical analysis was performed using Student's t-test with a significance level of P < 0.05. RESULT: TOF-SIMS analysis confirmed LPS contamination. LPS dentine affected SCAP viability but not adherence to dentine (P < 0.001). Conditioning of LPS dentine with either nanoparticles improved SCAP viability (P < 0.01) and rescued other LPS related adverse effects on SCAPs, such as F-actin disruption, decrease in differentiation/biomineralization potential. IL-6 produced by macrophages in response to LPS-treated dentine impeded SCAP migration (P < 0.001), diminished on CSnp and Dex-CSnp conditioning groups (P < 0.01). CONCLUSION: This study developed an LPS-dentine model and highlighted the ability of CSnp and Dex-CSnp to promote stem cell viability, migration, differentiation potential and reduce inflammation, providing an environment conducive for tissue regeneration/repair.

[Myxoid lipoblastoma:a clinicopathological study of four cases].

Objective: To study clinicopathological features,diagnosis and differential diagnosis of myxoid lipoblastoma. Methods: Four cases of myxoid lipoblastoma, from 2010 to 2017 at Wuxi People's Hospital of Nanjing Medical University, the Affiliated Hospital of Xuzhou Medical University and Binhai People's Hospital, were studied by clinicopathological analysis, immunohistochemistry and in situ hybridization along with a literature review. Results: The age of the patients ranged from 1 to 6 years. Histologically, all tumors had thin fibrous capsule and irregular lobules separated by fibrous septa. The individual lobules consisted of myxoid stroma,prominent plexiform capillary network and stellate or spindle mesenchymal cells. Lipoblasts (S-100 positive) and mature adipocytes varies among different lobules. FISH revealed PLAG1 disruption in all 4 cases. MDM2 or CHOP alterations were not detected. None of the patients had tumor recurrence upon follow up from 12 to 80 months. Conclusions: Myxoid lipoblastoma is a very rare tumor, usually in the first 5 years of life. The clinical features of myxoid lipoblastoma and lipoblastoma are similar, while myxoid lipoblastoma has prominent myxoid change, a plexiform vascular pattern and rare mature fat cells. The patient age,S-100 positive lipoblasts and cytogenetic alteration are the key diagnostic features.

Microtissue engineering root canal dentine with crosslinked biopolymeric nanoparticles for mechanical stabilization.

AIM: To evaluate the functional strain distribution pattern in root dentine following canal preparation and root canal surface engineering with crosslinked biopolymeric nanoparticles using digital moiré interferometry (DMI). METHODOLOGY: Root dentine specimens were prepared, grating material replicated and tested for 10-50 N, compressive loads in a customized high-resolution, whole-field moiré interferometry set-up. Digital moiré fringes were acquired to determine the strain distribution pattern at specific regions of interest before and after canal enlargement, and dentine surface engineering with a chitosan nanoparticle crosslinker solution. Fringe patterns were acquired, and strain distribution pattern in the direction perpendicular to dentinal tubules (U-field) and parallel to dentinal tubules (V-field) was analysed with custom digital image-processing software. Data were analysed with a statistical method on trend analysis at 0.05 significant level. RESULTS: Distinct deformation patterns perpendicular to the dentinal tubules were observed in root dentine. Root canal dentine removal following instrumentation resulted in an increase in strain distribution, which increased with an increase in applied loads (P < 0.01). The root canal dentine engineered with crosslinked nanoparticles demonstrated a conspicuous decrease in previously increased strain distribution in both coronal and apical root dentine (P < 0.01). A significant increase in tensile strain in root dentine was observed subsequent to instrumentation in the direction parallel to dentinal tubules (P < 0.01). There was a significant reduction in the tensile strain formed at the apical region of the instrumented root dentine following crosslinked nanoparticle treatment (P < 0.05). CONCLUSIONS: This study highlighted the potential of root canal dentine microtissue engineering with crosslinked chitosan nanoparticle to improve radicular strain distribution patterns in instrumented canals.

[Minimally invasive surgery for the severe degenerative lumbar scoliosis: two stage protocol].

Objective: To investigate the surgical outcome of minimally invasive surgery(MIS) for severe degenerative lumbar scoliosis(DLS) and put forward a two-stage MIS surgical strategy. Methods: Prospective study of MISDEF Ⅲ DLS patients from June 2016 to August 2017 in the Second Affiliated Hospital of Zhejiang University was carried out, excluding the patients whose apex vertebrae of scoliosis was above L(1/2) level or whose facet joint got spontaneous fusion. Fifty-three patients were included in this study for staging evaluation and MIS surgical treatment. Information was recorded, including gender, age, body mass index, follow-up period, pelvic incidence (PI), blood loss, operation time, visual analogue pain score (VAS), Oswestry disability index (ODI), complications in the perioperative period and follow-up period, and also the radiographic parameters such as scoliosis Cobb angle, the mismatch between pelvic incidence and lumbar lordosis (PI-LL), sagittalvertical axis (SVA), coronal balance (CB) before and after each stage of surgery or latest follow-up. The paired-samples t test was used to analyze the effectiveness of staging surgery. Results: Fifty-three patients (18 males and 35 females) were included in this study. All patients had completed clinical and the follow-up records, with an average follow-up period of 11.52 months (6-20 months). A total of 168 segments fusions were performed with CLIF, 113 segments were performed with anterior column realignment (ACR), the average correct angle was 15.6°±6.3°(7°-28°) in sagittal plane each level. After the stage Ⅰ surgery, lumbar scoliosis cobb had been corrected for 55.35%, after the stage Ⅱ surgery, rate of correction was 75.6%. PI-LL had been matched (-32.8°±14.9° to -2.5°±9.4°), SVA was changed from 5.7 cm to 0.6 cm, the stage Ⅰ rate of correction was 80.3 and stage Ⅱ was 88.8%, pelvic tilt (PT), lumbar lordosis (LL) and CB had been restored; 13 (24.5%) patients were performed paraspinal approach transforaminal decompression. The posterior minimally invasive fixation indexes: 11(20.8%) patients were performed paraspinal approach transforaminal multi-segment transforaminal osteotomy (TFO) and internal fixation; 36(67.92%) cases were performed paraspinal approach transforaminal multi-segment fixation; 6(11.33%) patients were treated with percutaneous pedicle screw fixation. The average fixed segments was 7.4±1.4 in each patient. The blood loss of stage Ⅰ and stage Ⅱ operation was (157±71) ml, (343±224)ml, respectively. The operation time was (214±60) min, (190±54)min respectively in the two stage operations. The low back pain and leg pain VAS score and ODI improved after the stage Ⅰ and Ⅱ surgery (t=17.948, 10.099, 14.619, all P<0.001). Conclusions: MIS for the severe degenerative lumbar scoliosis can achieve good clinical outcome and deformity correction. The two-stage protocol has the advantages of less complications and is well-tolerated.

[Application of autologous fat grafting in breast reconstruction].

Objective: To observe the outcome of breast reconstruction with autologous fat grafting in the patients following treatment for breast cancer. Methods: The clinical data of 22 patients after breast cancer modified radical mastectomy with fat grafting for breast reconstruction from January 2012 to March 2015 at Department of Body Contouring and Liposuction Center of Plastic Surgery, Hospital of Peking Union Medical College were analyzed retrospectively. The age of 22 patients (all female) was 28 to 54 years. Fifteen patients were performed breast modified radical mastectomy 5 to 16 year ago without radiotherapy, 7 patients were performed breast modified radical mastectomy following regular radiotherapy 2 years ago. Low negative pressure liposuction technical was applied to harvest fat tissue for 400 to 800 ml which was filtrated and purified by cotton pad method in low temperature environment. Fat grafting was performed with multi-level and multi-tunnel and in multi-point injection ways. All patients were followed up by regular imaging evaluation with MRI or ultrasonography after operation every 3 months. Results: All breast reconstruction were successfully performed in 22 patients, no severe complications occurred. Among 15 patients without radiotherapy, 12 patients were performed with autologous fat grafting for breast reconstruction, 3 patients with prosthetic implantation for breast augmentation after autologous fat grafting. Among 7 patients with radiotherapy, 6 patients were performed with autologous fat grafting for breast reconstruction, 1 patient with prosthetic implantation for breast augmentation after autologous fat grafting. The volume of fat grafting was 104 to 380 ml. It took 2.5 hours to finish the operation including 1.0 to 1.5 hours for liposuction and 40 minutes for fat grafting. Next fat grafting were performed after 3 months. The fat of the breast were survived well detecting by MRI, only 1 patient had a cystic nodule which had been resected during nipple reconstruction. Ultrasonography screened several cystic nodules with the major axis of 0.1 to 0.2 cm in the breast, which couldn't be found by palpating in 18 patients. The patients were followed up for 18 to 36 months, the outcome were satisfactory. Conclusion: Autologous fat grafting for breast reconstruction simplifies the operation program with satisfied results and avoids the complications of breast reconstruction with skin flap.

Effects of mutations on the structure and function of silkworm type 1 acetylcholinesterase.

AChE is the target of organophosphate (OP) and carbamate (CB) pesticides, and mutations in the gene can significantly reduce insects' sensitivity to these pesticides. Bombyx mori is highly sensitive to pesticides. To investigate the effects of mutations on AChE1 structure and function, we used a prokaryotic system to express B.mori wild type AChE1 (wAChE1) and mutant AChE1 (mAChE1) in this study. Active AChE1 proteins were obtained after refolding and purification, and wAChE1 and mAChE1 had similar activities. After incubation with 10(-6)M physostigmine and 10(-3)mg/mL phoxim, the remaining enzyme activity of mAChE1 was 4.42% and 8.86% higher than that of wAChE1's, respectively. Three-dimensional analysis of mutation AChE1 (mAChE1) revealed that the Ser and Ala side chains extended toward the central part of S285 with distances of just 2.80Å and 3.68Å, respectively, which changed the spatial structure of the active center and reduced its sensitivity to pesticides. These results indicated that the mutations altered the 3D structure of AChE1, which may affect the binding of physostigmine and phoxim to the serine residue at the active center, leading to reduced sensitivity. Our study helps understand the relationship between AChE1 mutations and pesticide resistance and provides a new direction for the cultivation of new pesticide-resistant varieties of B.mori.

Mechanisms of TiO2 NPs-induced phoxim metabolism in silkworm (Bombyx mori) fat body.

Silkworm is an important economic insect. Abuse of organophosphorus pesticides in recent years often leads to poisoning of silkworms, which significantly affects sericulture development by reducing silk production. Previous studies have shown that TiO2 NPs can effectively mitigate the damages caused by organophosphorus pesticides in silk glands and nerve tissues. The fat body is an important metabolic detoxification organ of silkworms, but it is unknown whether TiO2 NPs affect pesticide metabolism in fat body. In this study, we characterized the transcription of antioxidant genes and enzyme activity in fat body after TiO2 NPs and phoxim treatments using transcriptome sequencing, real-time PCR, and enzyme activity assay. Transcriptome sequencing detected 10 720, 10 641, 10 403, and 10 489 genes for control group, TiO2 NPs group, phoxim group, and TiO2 NPs+phoxim group, respectively. The TiO2 NPs+phoxim group had 705 genes with significantly differential expression (FDR<0.001), among which the antioxidant genes thioredoxin reductase 1 and glutathione S-transferase omega 3 were significantly upregulated. In phoxim group, the expression levels of superoxide dismutase (SOD), catalase (CAT), glutathione S-transferase delta (GSTd), and thioredoxin peroxidase (TPx) were increased by 1.365 -fold, 1.335 -fold, 1.642 -fold, and 1.765 -fold, respectively. The level changes of SOD, CAT, GSTd, and TPx were validated by real time PCR. The contents of reactive oxygen species (ROS), malondialdehyde (MDA), and hydrogen peroxide (H2O2) were increased by 1.598 -fold, 1.946 -fold, and 1.506 -fold, respectively, indicating that TiO2 NPs treatment can relieve phoxim-induced oxidative stress. To clarify the mechanism of TiO2 NPs's effect, the transcription levels of P450 gene family were measured for the TiO2 NPs+phoxim group; the expression levels of CYP4M5, CYP6AB4, CYP6A8, and CYP9G3 were elevated by 2.784 -fold, 3.047 -fold, 2.254 -fold, and 4.253 -fold, respectively, suggesting that high expression of P450 family genes can enhance the metabolism of phoxim in the fat body. The results of this study indicated that TiO2 NPs treatment promoted the transcriptional expression of the P450 family genes to improve the fat body's ability to metabolize phoxim and reduce phoxim-induced oxidative stress. This may be the main mechanism of TiO2 NPs' mitigation of phoxim-induced damages in the fat body.

Differentially expressed genes in the fat body of Bombyx mori in response to phoxim insecticide.

The silkworm, Bombyx mori, is an economically important insect. However, poisoning of silkworms by organophosphate pesticides causes tremendous loss to the sericulture. The fat body is the major tissue involved in detoxification and produces antimicrobial peptides and regulates hormones. In this study, a microarray system comprising 22,987 oligonucluotide 70-mer probes was employed to examine differentially expressed genes in the fat body of B. mori exposed to phoxim insecticide. The results showed that a total of 774 genes were differentially expressed upon phoxim exposure, including 500 up-regulated genes and 274 down-regulated genes. The expression levels of eight detoxification-related genes were up-regulated upon phoxim exposure, including six cytochrome P450s and two glutathione-S-transferases. It was firstly found that eight antimicrobial peptide genes were down-regulated, which might provide important references for studying the larvae of B. mori become more susceptible to microbial infections after phoxim treatment. In addition, we firstly detected the expression level of metamorphosis-related genes after phoxim exposure, which may lead to impacted reproduction. Our results may facilitate the overall understanding of the molecular mechanism of multiple pathways following exposure to phoxim insecticide in the fat body of B. mori.

Soil organic carbon stock and distribution in cultivated land converted to grassland in a subtropical region of China.

Land-use change from one type to another affects soil carbon (C) stocks which is associated with fluxes of CO2 to the atmosphere. The 10-years converted land selected from previously cultivated land in hilly areas of Sichuan, China was studied to understand the effects of land-use conversion on soil organic casrbon (SOC) sequestration under landscape position influences in a subtropical region of China. The SOC concentrations of the surface soil were greater (P\0.001) for converted soils than those for cultivated soils but lower (P\0.001) than those for original uncultivated soils. The SOC inventories (1.90­1.95 kg m-2) in the 0­15 cm surface soils were similar among upper, middle, and lower slope positions on the converted land, while the SOC inventories (1.41­1.65 kg m-2) in this soil layer tended to increase from upper to lower slope positions on the cultivated slope. On the whole, SOC inventories in this soil layer significantly increased following the conversion from cultivated land to grassland (P\0.001). In the upper slope positions, converted soils (especially in 0­5 cm surface soil) exhibited a higher C/N ratio than cultivated soils (P = 0.012), implying that strong SOC sequestration characteristics exist in upper slope areas where severe soil erosion occurred before land conversion. It is suggested that landscape position impacts on the SOC spatial distribution become insignificant after the conversion of cultivated land to grassland, which is conducive to the immobilization of organic C. We speculate that the conversion of cultivated land to grassland would markedly increase SOC stocks in soil and would especially improve the potential for SOC sequestration in the surface soil over a moderate period of time (10 years).

Characteristics of phoxim-exposed gene transcription in the silk gland of silkworms.

Silkworm (Bombyx mori), a model Lepidoptera insect, is an important economic insect. Its silk gland is the important organ for silk protein synthesis and secretion. Phoxim exposure causes deficient cocooning of silkworm and has become one of the major negative factors for the silk industry. To study the impact of phoxim exposure on silk gland, using gene chip technology, we examined differentially expressed genes in silk gland after silkworms were exposed to phoxim (4.0µg/mL) for 24h. Functional annotation, classification and KEGG signaling pathway analysis were performed. The results showed that out of 3206 genes detected in silk gland after phoxim exposure, 270 were differentially expressed significantly, including 249 up-regulated genes and 21 down-regulated genes. These differentially expressed genes related to apoptosis, detoxification and protein degradation were selected. Using qRT-PCR, the expression levels of 9 genes involved in apoptosis, detoxification and protein degradation were validated. In addition, the expression profiles of three related fibroin synthesis genes (Fib-H, Fib-L and P25) were analyzed. Our results showed that phoxim exposure induced apoptosis of silk gland cells and inhibition of fibroin synthesis. This may be the cause of deficient silkworm cocooning.

Effects of dietary electrolyte balance on growth performance, nitrogen metabolism and some blood biochemical parameters of growing rabbits.

The effects of different dietary electrolyte balance (DEB) on growth performance, nitrogen (N) metabolism and some blood biochemical parameters were investigated in 2 to 3 months old growing rabbits. A total of 150 growing rabbits of 2 months age were randomly divided into five groups according to average body weight, with 30 rabbits in each group. The DEB levels of the five experimental diets were -154, -3.16, +201, +347, and +500 meq/kg of dry matter (DM), respectively. There was a 7-d adaptation period and a 23-d experimental period. The results showed that the DEB levels had a quadratic affect on the average daily feed intake (ADFI) (p<0.001). The greatest ADFI was achieved when the DEB level was +201 meq/kg DM. Fecal N (FN) content linearly decreased (0.047), while digestible N (DN), retained N (RN), efficiency of intake N converted into digestible N (DN/IN) and the efficiency of intake N converted into retained N (RN/IN) linearly increased with the DEB increase (0.020, 0.004, 0.021, and 0.049, respectively). Serum phosphorus (P) ion content linearly increased with the DEB increase (p = 0.036). The DEB had a quadratic relationship with serum anion gap (AG) (p = 0.002) and serum parathyroid hormone (PTH) content (p = 0.016). The DEB levels quadratically affected base excess (BE) in the plasma (p<0.001). In conclusion, the DEB unaffected growth performance but affected feed intake, N metabolism and some blood biochemical parameters of growing rabbits.

Dual delivery for stem cell differentiation using dexamethasone and bFGF in/on polymeric microspheres as a cell carrier for nucleus pulposus regeneration.

This study aimed to investigate the feasibility of the nanostructured 3D poly(lactide-co-glycolide) (PLGA) constructs, which are loaded with dexamethasone (DEX) and growth factor embedded heparin/poly(L-lysine) nanoparticles via a layer-by-layer system, to serve as an effective scaffold for nucleus pulposus (NP) tissue engineering. Our results demonstrated that the microsphere constructs were capable of simultaneously releasing basic fibroblast growth factor and DEX with approximately zero order kinetics. The dual bead microspheres showed no cytotoxicity, and promoted the proliferation of the rat mesenchymal stem cells (rMSCs) by lactate dehydrogenase assay and CCK-8 assay. After 4 weeks of cultivation in vitro, the rMSCs-scaffold hybrids contained significantly higher levels of sulfated GAG/DNA and collagen type II than the control samples. Moreover, quantitative real time PCR analysis revealed that the expression of disc-matrix proteins including collagen type II, aggrecan, and versican in the rMSCs-scaffold hybrids was significantly higher than that in the control group, whereas the expression of osteogenic differentiation marker (collagen type I) was decreased. Taken together, these data indicate that Dex/bFGF PLGA microspheres could be used as a scaffold to improve the rMSCs growth and differentiating into NP like cells, and reduce the inflammatory response for IVD tissue engineering.

Growth performance, meat quality and Fatty Acid metabolism response of growing meat rabbits to dietary linoleic Acid.

An experiment was conducted to determine the effects of different amounts of dietary linoleic acid (LA) on growth performance, serum biochemical traits, meat quality, fatty acids composition of muscle and liver, acetyl-CoA carboxylase (ACC) and carnitine palmitoyl transferase 1 (CPT 1) mRNA expression in the liver of 9 wks old to 13 wks old growing meat rabbits. One hundred and fifty 9 wks old meat rabbits were allocated to individual cages and randomly divided into five groups. Animals in each group were fed with a diet with the following LA addition concentrations: 0, 3, 6, 9 and 12 g/kg diet (as-fed basis) and LA concentrations were 0.84, 1.21, 1.34, 1.61 and 1.80% in the diet, respectively. The results showed as follows: the dietary LA levels significantly affected muscle color of LL included a* and b* of experimental rabbits (p<0.05). The linear effect of LA on serum high density lipoprotein cholesterol was obtained (p = 0.0119). The saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs) contents of LL decreased and the polyunsaturated fatty acids (PUFAs) content of LL increased with dietary LA increase (p<0.0001). The PUFA n-6 content and PUFA n-3 content in the LL was significantly affected by the dietary LA levels (p<0.01, p<0.05). The MUFAs content in the liver decreased and the PUFAs contents in the liver increased with dietary LA increase (p<0.0001). The PUFA n-6 content and the PUFA n-6/n-3 ratio in the liver increased and PUFA n-3 content in the liver decreased with dietary LA increase (p<0.01). The linear effect of LA on CPT 1 mRNA expression in the liver was obtained (p = 0.0081). In summary, dietary LA addition had significant effects on liver and muscle fatty acid composition (increased PUFAs) of 9 wks old to 13 wks old growing meat rabbits, but had little effects on growth performance, meat physical traits and mRNA expression of liver relative enzyme of experimental rabbits.

Circle RNA circCSPP1 promotes human osteosarcoma cell proliferation and increases glucose metabolism by suppressing miR-200c maturation.

INTRODUCTION: MiR-200c plays a central role in glucose metabolism in cancer cells. However, its upstream regulators in this process are unknown. CircRNA CSPP1 (circCSPP1) was predicted to bind to premature miR-200c, an oncogenic miRNA. Therefore, we explored their interaction in osteosarcoma (OS). METHODS: Differential circCSPP1 and miR-200c expression in OS was analyzed using RT-qPCR. Glucose metabolism was analyzed by glucose uptake assay. Subcellular circCSPP1 location in OS cells was detected using cellular fractionation assay. The direct interaction between circCSPP1 and miR-200c was explored using RNA-RNA pull-down assay. The role of circCSPP1 in miR-200c maturation was investigated by analyzing both mature and premature miR-200c levels in OS cells with circCSPP1 overexpression. RESULTS: CircCSPP1 and premature miR-200c levels were increased while mature miR-200c level was decreased in OS. CircCSPP1 was detected in both the nuclear and cytoplasm fractions of OS cells. CircCSPP1 directly interacted with premature miR-200c. CircCSPP1 overexpression increased premature miR-200c level, glucose uptake, and cell proliferation, but decreased mature miR-200c level. MiR-200c overexpression suppressed the role of circCSPP1 in OS cells. CONCLUSIONS: CircCSPP1 promotes OS cell proliferation and increases glucose metabolism by suppressing miR-200c maturation.