Pulmonary Hypertension: Intensification and Personalization of Combination Rx (PHoenix): A phase IV randomized trial for the evaluation of dose-response and clinical efficacy of riociguat and selexipag using implanted technologies.

Approved therapies for the treatment of patients with pulmonary arterial hypertension (PAH) mediate pulmonary vascular vasodilatation by targeting distinct biological pathways. International guidelines recommend that patients with an inadequate response to dual therapy with a phosphodiesterase type-5 inhibitor (PDE5i) and endothelin receptor antagonist (ERA), are recommended to either intensify oral therapy by adding a selective prostacyclin receptor (IP) agonist (selexipag), or switching from PDE5i to a soluble guanylate-cyclase stimulator (sGCS; riociguat). The clinical equipoise between these therapeutic choices provides the opportunity for evaluation of individualized therapeutic effects. Traditionally, invasive/hospital-based investigations are required to comprehensively assess disease severity and demonstrate treatment benefits. Regulatory-approved, minimally invasive monitors enable equivalent measurements to be obtained while patients are at home. In this 2 × 2 randomized crossover trial, patients with PAH established on guideline-recommended dual therapy and implanted with CardioMEMS™ (a wireless pulmonary artery sensor) and ConfirmRx™ (an insertable cardiac rhythm monitor), will receive ERA + sGCS, or PDEi + ERA + IP agonist. The study will evaluate clinical efficacy via established clinical investigations and remote monitoring technologies, with remote data relayed through regulatory-approved online clinical portals. The primary aim will be the change in right ventricular systolic volume measured by magnetic resonance imaging (MRI) from baseline to maximal tolerated dose with each therapy. Using data from MRI and other outcomes, including hemodynamics, physical activity, physiological measurements, quality of life, and side effect reporting, we will determine whether remote technology facilitates early evaluation of clinical efficacy, and investigate intra-patient efficacy of the two treatment approaches.

[Analysis of surgical approaches and mandibular treatment strategies for different types of tumors involving parapharyngeal space].

In view of the surgical complexity of parapharyngeal space tumors involved, this paper summarized the disease data of patients with parapharyngeal space tumors involved in the Department of Oral and Maxillofacial Surgery, the First Hospital of Shanxi Medical University from January 2015 to January 2021. It also summarized the surgical approach and mandibular management, so as to explore surgical strategies for different characteristics of parapharyngeal space tumors involved. A total of 49 patients, including 28 males and 21 females, median age 52 years (range 24-72 years). They were treated with four surgical approaches for tumor resection, 25 cervical approach, 5 cheek and neck approach, 3 transoral approach, and 16 cervical-maxillary approach. Among the patients treated with cervical-maxillary approach, 3 patients were treated with mandible square resection, and 6 patients were treated with temporary mandible dissection. Seven cases were treated with tumor resection and partial mandibular resection. There are various surgical approaches and mandibular management methods involving tumors in the parapharyngeal space, and clinical decisions should be made based on tumor diameter, location, boundary, blood supply and pathological types.

[Genome sequence analysis of two SARS-CoV-2 virus infections in Inner Mongolia, 2022].

The target gene sequences of the novel coronaviruses obtained by sequencing were compared with the reference sequences to analyze the genetic variation of the two cases of the novel coronaviruses from Inner Mongolia Autonomous Region in 2022 and to explore the sources of infection. The results showed that the two sequences belonged to different evolutionary branches, Delta (AY.122) and Omicron (BA.1.1), respectively. hCoV-19/Inner Mongolia/IVDC-591/2022 had 48 single nucleotide polymorphisms on the genome sequences, sharing 40 nucleotide mutation sites with a Mongolian strain; hCoV-19/Inner Mongolia/IVDC-592/2022 genome shared 57 nucleotide mutation sites with a UK strain, and the nucleotide mutation site identity was 100% (57/57). Phylogenetic analysis showed that the target gene sequences were not directly related to domestic novel coronavirus sequences during the same period, but were related to isolates from Europe and Mongolia.

Inulin-induced differences on serum extracellular vesicles derived miRNAs in dairy cows suffering from subclinical mastitis.

MicroRNA (miRNA) profiles vary with the nutritional and pathological conditions of cattle. In this study, we aimed to investigate the effects of inulin supplement on miRNA profiles derived from serum extracellular vesicles (EVs). Our goal was to determine the differences in miRNA expressions and analyse the pathways in which they are involved. Based on the results of California mastitis test and milk somatic cell counts, ten lactating cows with subclinical mastitis were randomly divided into two groups: an inulin group and a control group (n = 5 in each group). The inulin group received a daily supplement of 300 g of inulin while the control group did not receive any supplementation. After a 5-week treatment period, serum-derived EV-miRNAs from each cow were isolated. High-throughput sequencing was conducted to identify differentially expressed miRNAs. GO and KEGG bioinformatics analysis was performed to examine the target genes of these differentially expressed miRNAs. The EV-RNA concentration and small RNA content were not affected by the inulin treatment. A total of 162 known miRNAs and 180 novel miRNAs were identified from 10 samples in the two groups. Among the known miRNAs, 23 miRNAs were found to be differentially expressed between the two groups, with 18 upregulated and five downregulated in the inulin group compared to the control group. Pathway analysis revealed the involvement of these differentially expressed miRNAs in the regulation of cell structure and function, lipid oxidation and metabolism, immunity and inflammation, as well as digestion and absorption of nutrients. Overall, our study provides a molecular-level explanation for the reported beneficial health effects of inulin supplementation in cows with subclinical mastitis.

Real-world effectiveness of pneumococcal vaccination in older adults: Cohort study using the UK Clinical Practice Research Datalink.

BACKGROUND: The 23-valent pneumococcal polysaccharide vaccine (PPV23) is recommended for UK older adults, but how age moderates effectiveness is unclear. METHODS: Three annual cohorts of primary-care patients aged≥65y from the Clinical Practice Research Datalink selected from 2003-5 created a natural experiment (n = 324,804), reflecting the staged introduction of the vaccine. The outcome was symptoms consistent with community-acquired pneumococcal pneumonia (CAP) requiring antibiotics or hospitalisation. We used the prior event rate ratio (PERR) approach to address bias from unmeasured confounders. RESULTS: Vaccinated patients had higher rates of CAP in the year before vaccination than their controls, indicating the potential for confounding bias. After adjustment for confounding using the prior event rate ratio (PERR) method, PPV23 was estimated to be effective against CAP for two years after vaccination in all age sub-groups with hazard ratios (95% confidence intervals) of 0.86 (0.80 to 0.93), 0.74 (0.65 to 0.85) and 0.65 (0.57 to 0.74) in patients aged 65-74, 75-79 and 80+ respectively in the 2005 cohort. Age moderated the effect of vaccination with predicted risk reductions of 8% at 65y and 29% at 80y. CONCLUSIONS: PPV23 is moderately effective at reducing CAP among UK patients aged≥65y, in the two years after vaccination. Vaccine effectiveness is maintained, and may increase, in the oldest age groups in step with increasing susceptibility to CAP.

[Progress in the methods for evaluating depth of bone invasion and bone invasion of lower gingival squamous cell carcinoma].

The depth of invasion is a new index in the 8th edition of TNM classification and staging of oral cancer. Currently, there is no standardized evaluation method for the diagnosis of bone invasion and depth of invasion in lower gingival squamous cell carcinoma (LGSCC). The evaluation of LGSCC bone invasion depth not only provides a reference for surgical margin, but also determines the choice of surgical method, and is an independent prognostic factor for predicting cervical lymphatic metastasis. At present, the main evaluation methods of LGSCC bone invasion and invasion depth include X-ray, MRI, CT, positron emission tomography(PET)/CT, PET/MRI, singlephoton emission CT(SPECT)/CT and pathological examination. In this paper, the evaluation methods and effects of LGSCC bone invasion and invasion depth are summarized, and its advantages and disadvantages are analyzed in order to provide reference for clinical application.

Blockage of PAK1 alleviates the proliferation and invasion of NSCLC cells via inhibiting ERK and AKT signaling activity

Purpose P21-activated kinase 1 (PAK1), a serine/threonine protein kinase which functions downstream of RAC and CDC42 GTPase, is activated by a variety of stimuli, including RAS and other growth signaling factors. The extracellular signal kinase (ERK) and protein kinase B (AKT) signal pathways have been implicated in the pathogenesis of cancers. Whether PAK1 is sensitive to KRAS mutation signals and plays a role through ERK and AKT signaling pathways in NSCLC needs to be studied. Methods The expression of PAK1, ERK and AKT was detected in both lung cancer cell lines and clinical samples. PAK1 RNA interference and specific inhibitor of PAK1(IPA-3) were applied to lung cancer cell lines and mouse xenograft tumors. Cell growth was measured by MTT and colony formation assays. Cell migration and invasion were detected by wound healing and transwell assays. RAS mutation was detected by Taqman probe method. Correlation between KRAS, PAK1, ERK and AKT activities was analyzed in lung cancer patients. Results PAK1 was highly expressed not only in RAS mutant but also in RAS wild-type lung cancer cells. Using specific inhibitor of PAK1, IPA-3 and PAK1 RNA interference, cell proliferation, migration and invasion of lung cancer cells were reduced significantly, accompanied by decreased activities of ERK and AKT. Dual inhibition of ERK and AKT suppressed these cellular processes to levels comparable to those achieved by reduction in PAK1 expression. In NSCLC patients, PAK1 was not correlated with KRAS mutation but was significantly positively correlated with pERK and pAKT. Conclusion PAK1 played roles in NSCLC proliferation and invasion via ERK and AKT signaling and suggested a therapeutic target for NSCLC (AU)

[Study on the effectiveness of silicon dust prevention and control of small sandblasting cabinet type water curtain exhaust hood].

Objective: To explore the effect of cabinet type water curtain exhaust hood applied to small sandblasting machine to prevent and control silicon dust, and put forward a new idea of dust ventilation protection facilities to effectively protect the occupational health of workers. Methods: From August to October 2018, the cabinet type water curtain exhaust hood of sandblasting room in a research institute was selected as the research object, and the methods of occupational health survey, on-site detection and physical simulation of air distribution were used to conduct on-site detection and smoke emission test on the local exhaust facilities, silica dust concentration, control wind speed and air distribution before and after the transformation line analysis and evaluation. Results: The air distribution simulation experiment showed that the air distribution of the cabinet type water curtain exhaust hood was reasonable and could effectively control the whole range of silica dust emission during the cleaning process. After modification, the capture velocity was increased from 0.01 m/s to 0.53 m/s, and the capture velocity was increased by 98.1%. The time weighted average allowable concentration (C(TWA)) of silicon dust (total dust) during sand blasting, cabin opening and cleaning was reduced from 7.00 mg/m(3) to 0.50 mg/m(3). The C(TWA) of silica dust (exhalation dust) was decreased from 3.36 mg/m(3) to 0.27 mg/m(3), and the C(TWA) dust reduction rates of total dust and respirable dust were 92.9% and 92.0%, respectively. Conclusion: The combination of cabinet type exhaust hood and water curtain dedusting optimizes the combination mode of dust prevention and control. It has the advantages of high efficiency of dedusting and purification, energy saving and environmental protection, and can be popularized and used in enterprises of the same nature.

Effects of propylene glycol on in vitro ruminal fermentation, methanogenesis, and microbial community structure.

We evaluated the effects of propylene glycol (PG) on in vitro ruminal fermentation, methanogenesis, and microbial community structure. A completely randomized design was conducted in the in vitro incubation, and 4 culture PG dose levels (0, 7.5, 15, and 22.5 µL/g of dry matter) were used in the trial. Based on the fermentation results, the control group (0 µL/g of dry matter, CON) and the second treatment group (15.0 µL/g of dry matter, TRT) were chosen for further analysis to explore the effects of PG on the bacterial and archaeal community structure. The concentrations of propanol, propanal, and succinate increased linearly, whereas the concentration of l-lactate decreased linearly as PG doses increased. The molar proportion of propionate demonstrated a linear increase with increasing PG doses. In contrast with propionate, the molar proportion of acetate and butyrate, and acetate-to-propionate ratio decreased linearly with increasing PG doses. The addition of PG markedly decreased methane production without negative effects on nutrient degradability. In the archaeal level, the relative abundance of Methanobrevibacter tended to decrease, but that of Methanomassiliicoccus significantly increased in TRT group. At the bacterial level, the relative abundance of Bacteroidetes and Prevotella in TRT group was numerically higher than that in CON group. The analysis of the Negativicutes class showed that the relative abundance of Succiniclasticum tended to increase, whereas that of Selenomonas tended to decrease in TRT group. These results demonstrated that PG might be used as an inhibitor to mitigate methane emission. However, the small decrease in methane production will limit the application of PG as a methane inhibitor in production practices. Further research is needed to determine whether use together with other inhibitors may improve the effects of PG on the utilization of reducing equivalents ([H]) and methane production.

Effect of different glucogenic to lipogenic nutrient ratios on rumen fermentation and bacterial community in vitro.

AIMS: This study was to investigate the effect of different ratios of glucogenic to lipogenic nutrients on rumen fermentation and the corresponding ruminal bacterial communities. METHODS AND RESULTS: Four diets, including glucogenic diet (G), lipogenic diet (L), two mixed diets: GL1 (G: L = 2 : 1) and GL2 (G:L = 1 : 2), served as substrates and were incubated with rumen fluid in vitro. The results revealed that the gas production, dry matter digestibility and propionate proportion were significantly increased by the G diet than others. The G diet increased the bacterial genera of Succinivibrionaceae_UCG_002, Succinivibrio, Selenomonas_1 and Ruminobacter but decreased some cellulolytic bacteria including the Eubacterium and several genera in family Ruminococcaceae than others. CONCLUSIONS: When the glucogenic nutrient was above 1/3 of the dietary energy source among the four diets, the in vitro incubation had a higher feed digestibility and lower acetate to propionate ratio. Bacterial genera, including Selenomonas, Succinivibrio, Ruminobacter, certain genera in Ruminococcaceae, Christensenellaceae_R-7_group and Eubacterium, were more sensitive to the glucogenic to lipogenic nutrients ratio. SIGNIFICANCE AND IMPACT OF THE STUDY: The present study provides a new perspective about the effect of dietary glucogenic to lipogenic ingredient ratios on rumen metabolism by comparing end-products, gas production and bacterial composition via an in vitro technique.

Blockage of PAK1 alleviates the proliferation and invasion of NSCLC cells via inhibiting ERK and AKT signaling activity.

PURPOSE: P21-activated kinase 1 (PAK1), a serine/threonine protein kinase which functions downstream of RAC and CDC42 GTPase, is activated by a variety of stimuli, including RAS and other growth signaling factors. The extracellular signal kinase (ERK) and protein kinase B (AKT) signal pathways have been implicated in the pathogenesis of cancers. Whether PAK1 is sensitive to KRAS mutation signals and plays a role through ERK and AKT signaling pathways in NSCLC needs to be studied. METHODS: The expression of PAK1, ERK and AKT was detected in both lung cancer cell lines and clinical samples. PAK1 RNA interference and specific inhibitor of PAK1(IPA-3) were applied to lung cancer cell lines and mouse xenograft tumors. Cell growth was measured by MTT and colony formation assays. Cell migration and invasion were detected by wound healing and transwell assays. RAS mutation was detected by Taqman probe method. Correlation between KRAS, PAK1, ERK and AKT activities was analyzed in lung cancer patients. RESULTS: PAK1 was highly expressed not only in RAS mutant but also in RAS wild-type lung cancer cells. Using specific inhibitor of PAK1, IPA-3 and PAK1 RNA interference, cell proliferation, migration and invasion of lung cancer cells were reduced significantly, accompanied by decreased activities of ERK and AKT. Dual inhibition of ERK and AKT suppressed these cellular processes to levels comparable to those achieved by reduction in PAK1 expression. In NSCLC patients, PAK1 was not correlated with KRAS mutation but was significantly positively correlated with pERK and pAKT. CONCLUSION: PAK1 played roles in NSCLC proliferation and invasion via ERK and AKT signaling and suggested a therapeutic target for NSCLC.

[Molecular mechanism of Puerariae Lobatae Radix in treatment of hepatocellular carcinoma based on network pharmacology].

To investigate the potential mechanism of Puerariae Lobatae Radix in the treatment of hepatocellular carcinoma by network pharmacology and in vitro cell experiment. The main active components of Puerariae Lobatae Radix and their predicted targets were obtained from TCMSP, and the disease targets were obtained from GeneCards database. The disease and drug prediction targets were intersected to select the common potential therapeutic targets. The "compound-target-disease" network diagram was constructed in Cytoscape 3.7.1, and the common targets were input into the STRING database to build the PPI network of proteins interaction. GO function and KEGG pathway enrichment analysis on effective targets were performed by using R software. Autodock vina 1.1.2 was used for molecular docking. Finally, the core targets and pathways were preliminarily verified by in vitro experiments. The proliferation of human hepatocellular carcinoma cells was detected by CCK-8 and EDU enzyme staining, and the expressions of PTEN, PDK1, Akt and GSK3 were detected by Western blot. In this study, 10 components of Puerariae Lobatae Radix(9 components involved in hepatocellular carcinoma-related targets and signaling pathways), and 149 hepatocellular carcinoma-related targets and 156 signaling pathways were screened out. The results of network analysis indicated that Puerariae Lobatae Radix may play an anti-hepatocellular carcinoma effect on key targets, such as Akt, IL6, MAPK3, EGFR, and key pathways, such as PI3 K-Akt. The results of molecular docking indicated that puerarin, genistein and daidzein had a good binding ability with the key targets such as AKT1, MAPK3, MAPK1 and CASP3, and puerarin had the lowest Vina score with AKT1 and MAPK3 and also similar to them. In vitro cell experiments confirmed that puerarin has a significantly inhibitory effect on the proliferation of human hepatocellular carcinoma cells. Western blot results showed that puerarin could increase the phosphorylation of PTEN in human hepatocellular carcinoma cells through the PTEN/Akt/GSK3ß signaling pathway, and the phosphorylation level of its downstream Akt decreased. This series of studies confirm that puerarin can treat hepatocellular carcinoma by blocking PTEN/Akt/GSK3ß cellular signaling pathway, so as to provide ideas for subsequent studies for the molecular mechanism of puerarin in the treatment of liver cancer.

[Correlation of interleukin-4 gene methylation regulation with the pathogenesis of recurrent aphthous ulcer].

Objetive: To investigate whether the methylation patterns of the interleukin-4 (IL-4) gene promoter changed and whether environmental factors affected the methylation level of IL-4 gene in the peripheral blood of patients with recurrent aphthous ulcer (RAU). Methods: Totally 20 patients, who were diagnosed with RAU, were recruited from May 2018 to May 2019 in the Department of Stomatology， First Hospital of Shanxi Medical University in the study (RAU group), including 12 females and 8 males, with mean age of 16-35 years. During the same period, 20 healthy volunteers matching the age and gender of the RAU group were selected from the medical personnel of the same hospital as the healty control group, including 11 females and 9 males, with mean age of 15-35 years. Peripheral blood samples of two groups were collected and the methylation levels of the IL-4 promoter were detected by bisulfite sequencing PCR (BSP). The IL-4 promoter methylation level of each sample was analyzed by direct sequencing and the IL-4 mRNA level was detected by real-time quantitative PCR. The data obtained were statistically analyzed. Results: The IL-4 gene promoter fragment contained 10 CPG sites from -1400 to -1625 bp. The methylation rates of CPG(-1556), CPG(-1483), CPG(-1479)and 10 CPG sites were significantly higher in RAU group ï¼»(32.0±19.9)%, (53.0±13.4)%, (46.0±19.8)% and (39.3±12.4)%ï¼½ than in healthy control group ï¼»(20.0±3.2)%, (35.5±12.3)%, (28.0±14.4)% and (32.6±5.8)%ï¼½, with statistically significant differences (P<0.05). The relative expression of IL-4 mRNA in the peripheral blood of RAU patients (1.0±0.1) was significantly lower than that of the healthy control group (1.5±0.2) (P<0.01). There was a significant negative correlation between the overall methylation rate of IL-4 gene promoter and the relative expression level of IL-4 mRNA in RAU group (r=-0.494, P<0.05). In the multivariate analysis, smoking, vitamin B12 and folic acid in the RAU group were significantly correlated with the overall methylation rate of the IL-4 gene promoter (P<0.01). Conclusions: The hypermethylation of IL-4 promoter in RAU patients may be related to the reduction of IL-4 gene transcription. Vitamin B12, folic acid and smoking may affect IL-4 gene methylation in peripheral blood of RAU patients.

Phenylboronic ester-modified anionic micelles for ROS-stimuli response in HeLa cell.

Smart polymers as ideal drug nanocarriers have attracted much attention due to the effective drug delivery, internalization and release once triggered by intracellular stimuli, as well as reduced cytotoxicity. We here reported the anionic micelle consisting of copolymer (PEG-b-PAsp) and a PBE (Phenylboronic Ester) group grafted, which can achieve fast response to intracellular ROS and enhanced anti-tumor activity. With this, PEG-b-PAsp-g-PBE/DOX system showed better tumor growth inhibition when studied on HeLa cell lines with high level of intracellular ROS and its subcutaneous tumor models. Additionally, the administration of PEG-b-PAsp-g-PBE/DOX did cause significantly lower systemic toxicity in comparison with free DOX. Hence, PEG-b-PAsp-g-PBE could be a highly efficient and safe nanocarrier to improve the efficacy of chemotherapeutic.

[Emerging of Japanese encephalitis virus and Getah virus from specimen of mosquitoes in Inner Mongolia Autonomous Region].

Objective: To investigate the types and distribution of blood-sucking insects and arboviruses in Inner Mongolia autonomous region, and provide basic data for the prevention of arbovirus transmitted disease. Methods: Blood-sucking insects were collected by lamp trapping method in nature. Mosquito samples were classified according to morphologic characteristics and then stored at liquid nitrogen. Viruses were isolated in cell culture and characterized, using molecular biological methods. Results: A total of 24 240 mosquitoes and 17 110 aphids were collected from 2 sites of 5 counties (Flags) in Inner Mongolia in 2014 and during 2017-2018. Among them, Japanese encephalitis virus gene was detected in Culex pipiens pallens, and 4 virus strains isolates which could be stably passaged. The isolates were identified as Getah virus and densonucleosis virus by molecular biology identification. Phylogenetic analysis on the E2 gene of the Getah virus (NMDK1813-1) showed that it belonged to the same evolutionary branch of the Gansu isolates (GS10-2) and having six common amino acid variation sites. Conclusions: The emergence of Japanese encephalitis virus and Getah virus from specimen of mosquitoes in Inner Mongolia indicated the new challenges on the prevention and control of arbovirus and related diseases. The results pf this study provided basic data for the prevention and control stretagies of arbovirus transmitted diseases in Inner Mongolia.

[Research progress in the methods for evaluating depth of invasion of tongue squamous cell carcinoma].

Depth of invasion is a newly added index for TNM staging of oral cancer in the eighth edition. Preoperative evaluation of depth of invasion not only provides a reference for surgical margin, but also serves as an independent prognostic factor for predicting lymph node metastases. At present, the main methods for assessing the depth of invasion of tongue squamous cell carcinoma include ultrasound examination, MRI, CT, positron emission tomography (PET) and histopathological examination. This paper summarizes the evaluation method and clinical effect of depth of invasion of tongue cancer, and analyzes its advantages and boundedness. In addition, this study is expected to provide a reference for the surgical treatment of tongue squamous cell carcinoma.

Effects of dietary threonine and immune stress on growth performance, carcass trait, serum immune parameters, and intestinal muc2 and NF-κb gene expression in Pekin ducks from hatch to 21 days.

An experiment was conducted to investigate the effects of different dietary threonine (Thr) levels and immune stress on Pekin ducklings' growth performance, carcass traits, serum immune parameters, and intestinal mucin 2 (MUC2) and nuclear factor kB (NF-κB) gene expressions. A total of 320 Pekin ducklings was randomly assigned to a 5 × 2 factorial arrangement of treatments. Each treatment group consisted of 4 replicate pens with 8 ducks per pen. Ducklings were fed 5 graded levels of Thr: 0.49, 0.56, 0.60, 0.65, and 0.76% from hatch to 21 d of age. At 11 d of age, ducks in the stressed groups were challenged with bovine serum albumin (BSA), and ducks in the unstressed groups were injected with normal saline water. The results showed that increasing Thr supplementation from 0.49 to 0.56% in the diet can improve BWG; feed consumption; weight and relative weight of breast and leg; weight of liver, bursa of Fabricius, spleen, and thymus; serum natural immune globulin A (IgA) concentration; and MUC2 gene expression in the ileum of 21-day-old Pekin ducks, significantly (P < 0.05). Immune stress with BSA had a significant effect on 21-day-old Pekin ducklings' BWG, feed consumption, and weight and relative weight of breast and thymus (P < 0.05), but no interaction between BSA and dietary Thr content was noticed in our experiment in 21-day-old Pekin ducks (P < 0.05). Dietary Thr requirements of the unstressed groups and stressed groups based on broken-line model analyses for ducks' BWG were 0.705 and 0.603%, respectively, and for ducks' feed consumption were 0.724 and 0.705%, respectively.

Clinical phenotype and outcome of hepatitis E virus-associated neuralgic amyotrophy.

OBJECTIVE: To determine the clinical phenotype and outcome in hepatitis E virus-associated neuralgic amyotrophy (HEV-NA). METHODS: Cases of NA were identified in 11 centers from 7 European countries, with retrospective analysis of demographics, clinical/laboratory findings, and treatment and outcome. Cases of HEV-NA were compared with NA cases without evidence of HEV infection. RESULTS: Fifty-seven cases of HEV-NA and 61 NA cases without HEV were studied. Fifty-six of 57 HEV-NA cases were anti-HEV IgM positive; 53/57 were IgG positive. In 38 cases, HEV RNA was recovered from the serum and in 1 from the CSF (all genotype 3). Fifty-one of 57 HEV-NA cases were anicteric; median alanine aminotransferase 259 IU/L (range 12-2,961 IU/L); in 6 cases, liver function tests were normal. HEV-NA cases were more likely to have bilateral involvement (80.0% vs 8.6%, p < 0.001), damage outside the brachial plexus (58.5% vs 10.5%, p < 0.01), including phrenic nerve and lumbosacral plexus injury (25.0% vs 3.5%, p = 0.01, and 26.4% vs 7.0%, p = 0.001), reduced reflexes (p = 0.03), sensory symptoms (p = 0.04) with more extensive damage to the brachial plexus. There was no difference in outcome between the 2 groups at 12 months. CONCLUSIONS: Patients with HEV-NA are usually anicteric and have a distinct clinical phenotype, with predominately bilateral asymmetrical involvement of, and more extensive damage to, the brachial plexus. Involvement outside the brachial plexus is more common in HEV-NA. The relationship between HEV and NA is likely to be causal, but is easily overlooked. Patients presenting with NA should be tested for HEV, irrespective of liver function test results. Prospective treatment/outcome studies of HEV-NA are warranted.

[Mononuclear cells of umbilical cord blood differentiation to granulocyte cell in vitro].

Objective: To explore an optimal method for granulocyte cell production from umbilical cord blood mononuclear cells. Methods: Erythrocytes were precipitated by hydroxyethyl starch. Mononuclear cells were isolated through Ficoll density gradient centrifugation. Different media, additives and cultivation model were chosen for granulocyte induction. Cell morphology was observed by microscopy, and cell phenotype was detected by flow cytometry. The CD18 expression of granulocytes was tested by immunofluorescence assay, and phagocytosis test was executed as well. Results: Compared to fetal bovine serum (FBS) treatment group, cell viability, counts and differentiation rate of granulocytes induced by X-VIVO(TM) 15 combined with TPO, SCF, G-CSF but without FBS were superior. And X-VIVO(TM)15 medium was better than SCGM medium at effectiveness and cost. Using two-stage mode of hematopoietic stem cell expansion followed by granulocyte induction with X-VIVO(TM)15 combining TPO, SCF and G-CSF, cell proliferation was nearly 132 times at day 21. Flow cytometry showed that the differentiation was lagged in 2-stage mode than in direct induction mode, CD15 expression was (69.60± 1.06) % vs (97.73±0.39) %; Wright-Giemsa staining demonstrated mature granulocytes; immunofluorescence showed the expression of lysosomal proteins CD18. A strong phagocytic function of mature granulocytes was demonstrated by phagotrophic efficiency of (51.43±0.05) %. And granulocyte had chemotaxis ability under the role of chemotactic factor IL-8. Conclusion: Optimized culture media and cultivation mode are achieved for functional granulocytes induction in vitro.

The effects of heat stress on protein metabolism in lactating Holstein cows.

Heat stress (HS) decreases milk protein synthesis beyond what would be expected based on the concomitant reduction in feed intake. The aim of the present study was to evaluate the direct effects of HS on milk protein production. Four multiparous, lactating Holstein cows (101 ± 10 d in milk, 574 ± 36 kg of body weight, 38 ± 2 kg of milk/d) were individually housed in environmental chambers and randomly allocated to 1 of 2 groups in a crossover design. The study was divided into 2 periods with 2 identical experimental phases (control phase and trial phase) within each period. During phase 1 or control phase (9 d), all cows were housed in thermal neutral conditions (TN; 20°C, 55% humidity) and fed ad libitum. During phase 2 or treatment phase (9 d), group 1 was exposed to cyclical HS conditions (32 to 36°C, 40% humidity) and fed ad libitum, whereas group 2 remained in TN conditions but was pair-fed (PFTN) to their HS counterparts to eliminate the confounding effects of dissimilar feed intake. After a 30-d washout period in TN conditions, the study was repeated (period 2), inverting the environmental treatments of the groups relative to period 1: group 2 was exposed to HS and group 1 to PFTN conditions. Compared with PFTN conditions, HS decreased milk yield (17.0%), milk protein (4.1%), milk protein yield (19%), 4% fat-corrected milk (23%), and fat yield (19%). Apparent digestibility of dry matter, organic matter, neutral detergent fiber, acid detergent fiber, crude protein, and ether extract was increased (11.1-42.9%) in HS cows, as well as rumen liquor ammonia (before feeding 33.2%; after feeding 29.5%) and volatile fatty acid concentration (45.3%) before feeding. In addition, ruminal pH was reduced (9.5 and 6% before and after feeding, respectively) during HS. Heat stress decreased plasma free amino acids (AA; 17.1%) and tended to increase and increased blood, urine, and milk urea nitrogen (17.2, 243, and 24.5%, respectively). Further, HS cows had reduced plasma glucose (8%) and nonesterified fatty acid (39.8%) concentrations compared with PFTN controls. These data suggest that HS increases systemic AA utilization (e.g., decreased plasma AA and increased nitrogen excretion), a scenario that limits the AA supply to the mammary gland for milk protein synthesis. Furthermore, the increase in AA requirements during HS might represent the increased need for gluconeogenic precursors, as HS is thought to prioritize glucose utilization as a fuel at the expense of nonesterified fatty acids.