Effects of rumen-encapsulated methionine and lysine supplementation and low dietary protein on nitrogen efficiency and lactation performance of dairy cows.

Low crude protein (CP) diets might be fed to dairy cows without affecting productivity if the balance of absorbed AA were improved, which would decrease the environmental effect of dairy farms. The aim of this study was to investigate the effects of supplementing ruminally protected Lys (RPL) and Met (RPM) at 2 levels of dietary CP on nutrient intake, milk production, milk composition, milk N efficiency (MNE), and plasma concentrations of AA in lactating Holstein cows and to evaluate these effects against the predictions of the new NASEM (2021) model. Fifteen multiparous cows were used in a replicated 3 × 3 Latin square design with 21-d periods. The 3 treatments were (1) a high-protein (HP) basal diet containing 16.4% CP (metabolizable protein [MP] balance of -130 g/d; 95% of target values), (2) a medium-protein diet containing 15% CP plus RPL (60 g/cow per day) and RPM (25 g/cow per day; MPLM; MP balance of -314 g/d; 87% of target values), and (3) a low-protein diet containing 13.6% CP plus RPL (60 g/cow per day) and RPM (25 g/cow per day; LPLM; MP balance of -479 g/d; 80% of target values). Dry matter intake was less for cows fed MPLM and LPLM diets compared with those fed the HP diet. Compared with the HP diet, the intake of CP, neutral detergent fiber, acid detergent fiber, and organic matter, but not starch, was lower for cows fed MPLM and LPLM diets. Milk production and composition were not affected by MPLM or LPLM diets relative to the HP diet. Milk urea N concentrations were reduced for the MPLM and LPLM diets compared with the HP diet, indicating that providing a low-protein diet supplemented with rumen-protected AA led to greater N efficiency. There was no significant effect of treatment on plasma AA concentrations except for proline, which significantly increased for the MPLM treatment compared with the other 2 treatments. Overall, the results supported the concept that milk performance might be maintained when feeding lactating dairy cows with low CP diets if the absorbed AA balance is maintained through RPL and RPM feeding. Further investigations are needed to evaluate responses over a longer time period with consideration of all AA rather than on the more aggregated MP and the ratio between Lys and Met.

Hydroponic barley supplementation fed with high-protein diets improves the production performance of lactating dairy cows.

The study investigated the effects of dietary protein level and the inclusion of hydroponic barley sprouts (HB) on lactation performance, blood biochemistry and N use efficiency in mid-lactation dairy cows. Treatments were arranged in a 2 × 2 factorial design with 2 CP levels (16.8% and 15.5% of DM), with HB (4.8% of DM, replacing 4.3% of alfalfa hay and 0.5% of distillers dried grains with solubles [DDGS]) or without HB. Forty-eight multiparous Holstein dairy cows (146 ± 15 DIM, 40 ± 5 kg/d of milk) were randomly allocated to 1 of 4 diets: high-protein diet (16.8% CP, HP), HP diet with HB (HP+HB), low-protein diet (15.5% CP, LP), or LP diet with HB (LP+HB). An interaction between CP × HB on DMI was detected, with DMI being unaffected by HB inclusion in cows fed the high-protein diets, but was lower in cows fed HB when the low-protein diet was fed. A CP × HB interaction was also observed on milk and milk protein yield, which was higher in cows fed HB with HP, but not LP. Inclusion of HB also tended to reduce milk fat content, and feeding HP resulted in a higher milk protein and MUN content, but lower milk lactose content. Feed efficiency was increased by feeding HP or HB diets, whereas N use efficiency was higher for cows fed LP or HB diets. There was an interaction on the apparent total-tract digestibility of DM and CP, which was higher when HB was fed along with HP, but reduced when fed with LP, whereas the digestibility of ADF was increased by feeding low-protein diets. In conclusion, feeding a low-protein diet had no adverse effect on cow performance, while feeding HB improved milk and milk component yield, and N efficiency when fed with a high-CP diet, but compromised cow performance with a low-CP diet.

[A successfully treated case of respiratory failure caused by eating Nassariidaes].

Food poisoning caused by Nassariidaes has occurred frequently in coastal areas of China, especially in summer and autumn. Nassariidaes poisoning can be manifested as lip and tongue paralysis, dizziness, headache, nausea and vomiting, arrhythmia and even respiratory failure. We admitted a case of respiratory failure caused by eating Nassariidaes. After timely respiratory support, hemoperfusion and other active treatment, the patient was recovered and was discharged. This paper summarized clinical characteristics and treatment of Nassariidaes poisoning, in order to provide reference for clinical diagnosis and treatment of similar cases.

[Progress in clinical research on potential therapeutic drugs for acute-on-chronic liver failure].

Acute-on-chronic liver failure (ACLF), has a high mortality rate and a poor prognosis. Currently, the only effective treatment for ACLF is liver transplantation. However, the number of patients who can successfully undergo liver transplantation is limited due to the rapid progression of ACLF, the occurrence of serious complications, and a dearth of liver donors. The available drug treatment indication expansion and pathogenesis exploration are expected to delay the progression of ACLF, reduce complications, and provide patients with opportunities for liver transplantation by improving portal vein pressure, inhibiting excessive inflammatory response, correcting energy metabolism disorders, reducing oxidative stress, resisting hepatic cell apoptosis, and promoting liver regeneration.

[Effects of CACNA1H gene knockout on autistic-like behaviors and the morphology of hippocampal neurons in mice].

OBJECTIVE: To investigate the effects of CACNA1H gene knockout (KO) on autistic-like behaviors and the morphology of hippocampal neurons in mice. METHODS: In the study, 25 CACNA1H KO mice of 3-4 weeks old and C57BL/6 background were recruited as the experimental group, and 26 wild type (WT) mice of the same age and background were recruited as the control group. Three-chamber test and open field test were used to observe the social interaction, anxiety, and repetitive behaviors in mice. After that, their brain weight and size were measured, and the number of hippocampal neurons were observed by Nissl staining. Furthermore, the CACNA1H heterozygote mice were interbred with Thy1-GFP-O mice to generate CACNA1H-/--Thy1+(KO-GFP) and CACNA1H+/+-Thy1+ (WT-GFP) mice. The density and maturity of dendritic spines of hippocampal neurons were observed. RESULTS: In the sociability test session of the three-chamber test, the KO mice spent more time in the chamber of the stranger mice than in the object one (F1, 14=95.086, P < 0.05; Post-Hoc: P < 0.05), without any significant difference for the explored preference index between the two groups (t=1.044, P>0.05). However, in the social novelty recognition test session, no difference was observed between the time of the KO mice spend in the chamber of new stranger mice and the stranger one (F1, 14=18.062, P < 0.05; Post-Hoc: P>0.05), and the explored preference index of the KO mice was less than that of the control group (t=2.390, P < 0.05). In the open field test, the KO mice spent less time in the center of the open field apparatus than the control group (t=2.503, P < 0.05), but the self-grooming time was significantly increased compared with the control group (t=-2.299, P < 0.05). Morphological results showed that the brain weight/body weight ratio (t=0.356, P>0.05) and brain size (t=-0.660, P>0.05) of the KO mice were not significantly different from those of the control group, but the number of neurons were significantly reduced in hippocampal dentate gyrus compared with the control group (t=2.323, P < 0.05). Moreover, the density of dendritic spine of dentate gyrus neurons in the KO-GFP mice was significantly increased compared with the control group (t=-2.374, P < 0.05), without any significant difference in spine maturity (t=-1.935, P>0.05). CONCLUSION: CACNA1H KO mice represent autistic-like behavior, which may be related to the decrease in the number of neurons and the increase in the density of dendritic spine in the dentate gyrus.

[Predictive value of impedance of leadless pacemaker during implantation on trend changes of pacing threshold].

Objective: To explore the predictive value of the impedance measured during leadless pacemaker Micra implantation on the trend of changes of pacing threshold post implantation. Methods: This is a retrospective cross-sectional study. Patients who received implantation of leadless pacemaker Micra at the Second Xiangya Hospital of Central South University from December 2019 to August 2020 were enrolled. The clinical data and the intraoperative electrical parameters during leadless pacemaker implantation were collected. The impedance and pacing threshold data were analyzed at three time points: immediate release, 5-10 min after release, and after traction test. Receiver operating characteristic (ROC) curves and the area under the curve (AUC) were used to analyze the value of the impedance at immediate release on predicting the trend of changes of pacing threshold post implantation. Results: A total of 21 patients (mean age: (72.2±12.5) years, 12 males) were included. The impedance of 21 patients was (798.1±35.3) Ω immediately after implantation, (800.9±35.6) Ω after 5-10 minutes of release, and (883.6±31.7) Ω after traction test. Impedance was similar between the three time points (P>0.05). The threshold was (0.97±0.11) V/0.24 ms immediately after implantation, (0.95±0.12) V/0.24 ms at 5-10 min after the release, and (0.59±0.06) V/0.24 ms after the traction test. The threshold was significantly lower after the traction test than that immediately after release (P=0.003) and than that at 5-10 minutes after release (P=0.008), suggesting a decreased tendency of the threshold over time. According to the analysis of the ROC curve, the immediate impedance after the release ≥680 Ω could predict the ideal pacing threshold after the traction test (AUC=0.989, 95%CI 0.702-0.964, P<0.001), the prediction sensitivity was 87%, and the specificity was 100%. The pacing threshold would be not ideal with the immediate impedance ≤ 520 Ω (95%CI 0.893-1.000, P<0.001), the sensitivity was 100%, and the specificity was 80%. Conclusions: The impedance immediately after the release has predictive value for the changing trend of threshold post leadless pacemaker Micra implantation. Impedance ≥680 Ω immediately after release is often related with ideal pacing threshold after the traction test. In contrast, the impedance ≤ 520 Ω pacing is often related with unsatisfactory threshold after the traction test, therefore, it is recommended to find a new pacing site to achieve the impedance ≥680 Ω immediately after release during leadless pacemaker Micra implantation.

[Establishment of reference interval of vitamin A for Chinese children and adolescents based on expectation-maximization algorithm].

Objective: To establish the reference interval of vitamin A for Chinese children and adolescents by indirect approach. Method: The serum retinol samples from 28 provinces, autonomous regions and municipalities in China between January 2016 and June 2019 were examined by high performance liquid chromatography in third-party clinical laboratory institutions. Using a cross-sectional study, descriptive statistics of serum vitamin A levels in children and adolescents aged 0-18 years were carried out in population, region and season. The distribution parameters of vitamin A levels were calculated iteratively by the expectation-maximization (EM) algorithm, and the reference interval was established. Results: A total of 1 096 597 cases were included in the analysis, including 631 017 males and 465 580 females, with a median age of 3(1, 6) years. Before the age of 4 years, the median serum vitamin A level increased with age [median level and its quartiles was 0.15 (0.11, 0.18) mg/L for neonates, 0.23 (0.18, 0.29) mg/L for infants aged below 1 year, and 0.27 (0.20, 0.34) mg/L for kids aged 1-<4 years]. The median serum vitamin A level was stable at the age of 4-8 years (median level ranged from 0.27 to 0.29 mg/L). Afterwards, the median serum vitamin A level reached 0.30 mg/L at the age of 9 years, and then an increasing trend with age was observed till 18 years. The median serum vitamin A level was higher in south area (male:0.33 mg/L; female:0.34 mg/L), compared with other areas (0.24-0.30 mg/L). The variation of the median serum vitamin A level among seasons was within 0.03 mg/L. Based on EM algorithm using indirect approach, the reference intervals of vitamin A for Chinese children and adolescents were as follows: newborn, 0.27-0.47 mg/L; infants aged below 1 year, 0.36-0.56 mg/L; children aged 1-<7 years, 0.35-0.54 mg/L; children aged 7-<16 years, 0.46-0.60 mg/L; and teenagers aged over 16 years, 0.38-0.59 mg/L. Conclusion: The EM algorithm can provide a basis for the establishment of vitamin A reference interval for Chinese children and adolescents, and the established reference interval needs to be further verified by direct method.

[Treatment report of three cases of multiple wasp stings].

From September to November 2019, three patients with Hornet stings were admitted to hospital with acute anaphylaxis and various degrees of organ dysfunction. After hemoperfusion combined with continuous hemodialysis and hormone pulse therapy, 1 patient was successfully treated and 2 patients were not cured. Bee Stings can cause systemic allergic reactions and toxic reactions. In severe cases, acute anaphylaxis, rhabdomyolysis, intravascular hemolysis, abnormal coagulation, and acute liver and kidney injury may occur, myocardial damage and other multi-organ function damage or even failure, is a clinical acute and critical illness, mortality is higher. Through the analysis of the diagnosis and treatment process of these 3 cases, the experience is summarized: the Bee sting should be treated as soon as possible, the early prevention and treatment of multiple organ dysfunction, the choice of accurate blood purification and comprehensive treatment, can effectively improve the prognosis, increase the success rate of treatment.

[Epidemic characteristics of hand, foot and mouth disease (HFMD) and the lag effect of average daily temperature on the epidemic of HFMD in Beijing].

Objective: To analyze the epidemic characteristics of hand, foot and mouth disease (HFMD) in Beijing and explore the short-term lag effect of daily average temperature on HFMD. Methods: The incidence data and meteorological data of HFMD were collected from Xicheng District, Chaoyang District, and Changping District of Beijing between January 2009 and December 2018. The dose-response relationship and short-term lag effect of temperature and HFMD incidence were explored by the distributed lag non-linear model (DNLM). Results: From 2009 to 2018, a total of 97 210 cases of HFMD were confirmed in Xicheng District, Chaoyang District, and Changping District of Beijing. The incidence rate of HFMD in Xicheng district was 71.83/100 000, which was the lowest in three districts. The total annual average incidence was 146.89/100 000 in the three districts. The cases were concentrated from May to July, with a small peak in October to November. The ratio of male to female was 1.49∶1, and the median age of the patients was 3.08 (1.89, 4.39) years old, which showed a decreasing trend in children under 6 years old (Z=-30.11, P<0.01). The DNLM showed that the cumulative relative risk (RR) curve had a bimodal distribution, with RR of 1.36 (95%CI: 1.05-1.76) and 1.35 (95%CI: 1.10-1.66), and the peak values appeared at 4 ℃ and 26 ℃, respectively. Conclusions: The incidence rate of HFMD in Xicheng was the lowest in three districts of Beijing. In addition, the incidence of HFMD is seasonal, and the incidence is higher in spring and summer than that in autumn and winter. Daily average temperature had a lag effect on HFMD, which was different between high temperature and low temperature.

Identification of downstream target genes regulated by CX43 in hepatocellular carcinoma.

The aim of study was to identify the downstream target genes of CX43 by Human Transcriptome Array. Therefore, a gene microarray was generated which consists of CX43-overexpressed hepatocellular carcinoma (HCC) cells transfected with the constructed plasmid and negative controls to identify candidate genes. Integrated bioinformatic analysis was used to clarify biological functions of the identified genes, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, protein-protein interaction network, and survival analysis. The candidate genes were further validated by qRT-PCR in liver cancer tissues and CX43-silenced HCC cells. We have found the mRNA and protein levels of CX43 significantly upregulated in HCC cells transfected with CX43 constructed plasmid. We identified 928 differentially expressed genes including 394 upregulated and 534 downregulated genes, enriched in the cancer related functions and pathways by GO and KEGG pathway analysis. The protein-protein interaction network revealed 9 hub genes in this study. Statistical analysis indicated that upregulation of RALA and SRC was associated with poor prognosis in liver cancer. The differential expression of 2 candidate genes were further validated in HCC cells and tissues. In conclusion, protein-coding genes RALA and SRC could be target genes of CX43 and therapeutic targets for hepatocellular carcinoma.

Effects of antibiotic residues in milk on growth, ruminal fermentation, and microbial community of preweaning dairy calves.

The aim of this study was to evaluate the effects of antibiotic residues in milk on growth, ruminal fermentation, and microbial community of dairy calves in their first 35 d of age. Twenty newborn Holstein bull calves were assigned to 1 of 2 treatments equally: milk replacer without antibiotics (control) and milk replacer plus 4 antibiotics: 0.024 mg/L of penicillin, 0.025 mg/L of streptomycin, 0.1 mg/L of tetracycline, and 0.33 mg/L of ceftiofur (ANT). Starter intake and fecal consistency scores of each calf were recorded on a daily basis. Body weight, withers height, body length, and heart girth were measured on d 1, 7, 14, 21, 28, and 35 before feeding in the morning. Rumen fluid was collected on d 15, 25, and 35 to determine ruminal pH, volatile fatty acids (VFA), and NH3-N concentrations. A total of 10 (5 per treatment) samples of rumen fluid taken on d 35 were analyzed for microbial community. Rumen tissues from the cranial ventral sac and cranial dorsal sac were collected from 8 calves of each group for morphology analysis on d 35 after being harvested. The results showed that calves in 2 treatments had similar starter intake, body weight, withers height, body length, heart girth, and average daily gain. The ANT group showed a lower diarrhea frequency in wk 4, and no differences were found for other weeks. Calves in the ANT group exhibited a greater concentration of acetic acid in the rumen and no differences for other VFA, total VFA, rumen pH, or NH3-N. As for rumen morphology, the length of papillae from cranial ventral sac of the ANT group was longer than that of the control group. The results of ruminal microbial community showed that antibiotic residues had minor effects on bacteria phyla and bacteria diversity. At the genus level, calves in the ANT group showed lower richness of Prevotella and higher richness of Acetitomaculum. In conclusion, antibiotic residues stimulated the development of ruminal papillae and increased the production of acetic acid in rumen, which might be caused by the influence of antibiotics on the ruminal microbial community.

[The clinical application of ventriculovesical shunts].

Objective: To report a clinical protocol for the application of ventriculovesical shunts (VV shunts) in patient who may be unable to maintain or receive ventriculoperitoneal (VP) shunts. Methods: 25 patients underwent placement of VV shunts as an alternative to VP shunt therapy from March 2010 through March 2016. They all have malfunction of the VP shunt for the following reasons: peritoneal fluid inclusion(11 cases), failure of the peritoneum to absorb cerebrospinal fluid(7 cases), intra-abdominal infection(5 cases), peritoneal irritation pain(2 cases). The patients ranged in age from 5 years to 68 years old. All the patients underwent preoperative examine including urine routines, renal function electrolyte examination, imaging of the urinary bladder by ultrasonography. Results: Two patients died from other diseases, one patient's shunts were removed due to proximal catheter occlusion 2 years after the operation, the penis pain was occurred in one patient. The rest patients had functional VV shunts. Conclusions: Ventriculovesical shunts may be considered for the treatment of hydrocephalus when the peritoneal cavity cannot be used as a distal terminus.

[A prognostic scoring scale for adult patients with supratentorial primary anaplastic gliomas].

Objective: This study explored the prognostic factors of patients with supratentorial anaplastic gliomas and tried to propose a prognostic scoring scale with aim to provide theoretical reference for clinical treatment. Methods: The clinical data of 198 patients surgically treated for primary anaplastic glioma in Henan Provincial People's Hospital between Jan 2009 and Jan 2018 were reviewed. Univariate and multivariate analyses were used to identify prognostic factors with methods of Kaplan-Meier plot and Cox proportional hazard model, respectively. Based on the prognostic factors, a scoring scale was thereby proposed. Results: Univariate analysis results showed age, tumor location, tumor diameter, preoperative KPS, extent of resection, radiotherapy, chemotherapy, pathology with oligodendroglial components, 1p/19q, IDH, MGMT were significantly correlated with survival (P<0.05). Multivariate analysis results showed age ≥45 years old, tumor diameter ≥6 cm, preoperative KPS<70, without radiotherapy, 1p/19q intact, MGMT promoter unmethylation were independent prognostic risk factors (P<0.05). Patients were scored with 0-6 points based on the formulation that each independent prognostic risk factor was assigned with 1 point. Then patients were further grouped according to the score. Those with less than 2 points were low-risk group, equal to 2 points were medium-risk group, equal to 3 points were high-risk group, more than or equal to 4 points were extremely high-risk group. There were significant differences in survival between the different groups (P<0.000 1). Conclusions: The higher score, the shorter survival time. This prognostic scoring scale can provide a theoretical basis for the prognosis estimation of patients with anaplastic glioma and help to carry out personalized clinical treatment.

Tuning the electrical and optical anisotropy of a monolayer black phosphorus magnetic superlattice.

We investigate theoretically the effects of modulated periodic perpendicular magnetic fields on the electronic states and optical absorption spectrum in monolayer black phosphorus (phosphorene). We demonstrate that different phosphorene magnetic superlattice (PMS) orientations can give rise to distinct energy spectra, i.e. tuning the intrinsic electronic anisotropy. Rashba spin-orbit coupling (RSOC) develops a spin-splitting energy dispersion in this phosphorene magnetic superlattice. Anisotropic momentum-dependent carrier distributions along/perpendicular to the magnetic strips are demonstrated. The manipulations of these exotic electronic properties by tuning superlattice geometry, magnetic field and the RSOC term are addressed systematically. Accordingly, we find bright-to-dark transitions in the ground-state electron-hole pair transition rate spectrum and the PMS orientation-dependent anisotropic optical absorption spectrum. This feature offers us a practical way of modulating the electronic anisotropy in phosphorene by magnetic superlattice configurations and detecting this modulation capability by using an optical technique.

Nuclear factor erythroid 2-related factor 2 antioxidant response element pathways protect bovine mammary epithelial cells against H2O2-induced oxidative damage in vitro.

The experiment was conducted to determine the role of nuclear factor (erythroid-derived 2)-like factor 2 (NFE2L2, formerly Nrf2) antioxidant response element (ARE) pathway in protecting bovine mammary epithelial cells (BMEC) against H2O2-induced oxidative stress injury. An NFE2L2 small interfering RNA (siRNA) interference or a pCMV6-XL5-NFE2L2 plasmid fragment was transfected to independently downregulate or upregulate expression of NFE2L2. Isolated BMEC in triplicate were exposed to H2O2 (600 µM) for 6 h to induce oxidative stress before transient transfection with scrambled siRNA, NFE2L2-siRNA, pCMV6-XL5, and pCMV6-XL5-NFE2L2. Cell proliferation, apoptosis and necrosis rates, antioxidant enzyme activities, reactive oxygen species (ROS) and malondialdehyde (MDA) production, protein and mRNA expression of NFE2L2 and downstream target genes, and fluorescence activity of ARE were measured. The results revealed that compared with the control, BMEC transfected with NFE2L2-siRNA3 had proliferation rates that were 9 or 65% lower without or with H2O2, respectively. These cells also had apoptosis and necrosis rates that were 27 and 3.5 times greater with H2O2 compared with the control group, respectively. In contrast, transfected pCMV6-XL5-NFE2L2 had proliferation rates that were 64.3% greater or 17% lower without or with H2O2 compared with the control group, respectively. Apoptosis rates were 1.8 times lower with H2O2 compared with the control. In addition, compared with the control, production of ROS and MDA and activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and glutathione-S-transferase (GST) increased markedly in cells transfected with pCMV6-XL5-NFE2L2 and without H2O2. However, compared with the control, production of ROS and MDA and activity of CAT and GSH-Px increased markedly, whereas activities of SOD and GST decreased in cells transfected with pCMV6-XL5-NFE2L2 and incubated with H2O2. Compared with the control, cells transfected with NFE2L2-siRNA3 with or without H2O2 had lower production of ROS and MDA and activity of SOD, CAT, GSH-Px, and GST. Cells transfected with pCMV6-XL5-NFE2L2 with or without H2O2 had markedly higher protein and mRNA expression of NFE2L2, heme oxygenase-1 (HMOX-1), NADH quinone oxidoreductase 1, glutamate cysteine ligase catalytic subunit, and glutamyl cystine ligase modulatory subunit compared with the control incubations. Cells transfected with NFE2L2-siRNA3 without or with H2O2 had markedly lower protein and mRNA expression of NFE2L2, HMOX-1, NADH quinone oxidoreductase 1, glutamyl cystine ligase modulatory subunit, and glutamate-cysteine ligase catalytic subunit compared with the control incubations. In addition, expression of HMOX-1 was 5.3-fold greater with H2O2 compared with the control. Overall, results indicate that NFE2L2 plays an important role in the NFE2L2-ARE pathway via the control of HMOX-1. The relevant mechanisms in vivo merit further study.

Nuclear factor erythroid 2-related factor 2-antioxidant activation through the action of ataxia telangiectasia-mutated serine/threonine kinase is essential to counteract oxidative stress in bovine mammary epithelial cells.

Nuclear factor (erythroid-derived 2)-like factor 2 (NFE2L2, formerly Nrf2) is a transcription factor that binds to the antioxidant response element (ARE) in the upstream promoter region of various antioxidant-responsive genes. Hence, at least in nonruminants, the NFE2L2-ARE signaling pathway plays an important role in the cellular antioxidant defense system. Whether oxidative stress in bovine mammary epithelial cells alters NFE2L2 or the NFE2L2-ARE pathway is unclear. Therefore, the objective of this study was to examine the response in NFE2L2- and NFE2L2-ARE-related components in bovine mammary epithelial cells (BMEC) under oxidative stress. An in silico analysis to identify potential phosphorylation sites on NFE2L2 and the protein kinases was performed with Netphos 3.1 (http://www.cbs.dtu.dk/services/NetPhos/) and Scansite (http://scansite.mit.edu) software. Isolated BMEC were exposed to H2O2 (600 µM) for 6 h to induce oxidative stress. In silico analysis revealed ataxia telangiectasia-mutated (ATM) serine/threonine kinase as a key kinase responsible for the phosphorylation of NFE2L2. Thus, after the 6 h incubation with H2O2, BMEC were transiently transfected with ATM-small interfering RNA (siRNA) 1, 2, or 3. Compared with the control, transfection with ATM-siRNA3 resulted in proliferation rates that were 60.7 and 36.2% lower with or without H2O2. In addition, production of reactive oxygen species and malondialdehyde increased markedly, but activities of superoxide dismutase, glutathione peroxidase, catalase, and glutathione-S-transferase decreased markedly in transfected cells without or with H2O2 compared with the control. Transfected cells had markedly lower protein and mRNA expression of NFE2L2 without or with H2O2 compared with the control. In addition, fluorescent activity of the ARE in transfected BMEC indicated that NFE2L2-driven transcriptional activation decreased under oxidative stress. Overall, results indicate that ATM is a physiologically relevant NFE2L2 kinase. Furthermore, inhibition of ATM activity can cause marked alterations in oxidative stress leading to cell death as a result of diminished capacity of BMEC to cope with H2O2-induced cytotoxicity. The relevance of this kinase in vivo merits further study.

[Interventional treatment of symptomatic intracranial in-stent restenosis].

Objective: To assess the feasibility and safety of interventional treatment of symptomatic intracranial in-stent restenosis (SISR). Methods: Clinical data of 21 patients with SISR who underwent interventional treatment in the General Hospital of the PLA Rocket Force from January 2012 to May 2018 were retrospectively analyzed. Perioperative complications, angiographic and clinical follow-up results were recorded. Results: The success rate of treatment was 100%, including 21 lesions of SISR (7 at the V4 segment of the vertebral artery, 7 at the basilar artery, 5 at the M1 segment of middle cerebral artery, 2 at the internal carotid artery). The residual stenosis rate was preoperative 84±7, postoperative 30±14 respectively. Balloon angioplasty and stent implantation were performed in 10 patients (48.6%) and 11 patients (52.4%), respectively. After treatment, 1 patient experienced perforating event without neurological sequelae. Of the 11 patients (52.4%) completed angiography follow-up, 3 (3/11, 27.3%) ocurred restenosis and 1 was retreated. During clinical follow-up, 1 patients received intravenous thrombolysis for the symptom of acute cerebral infarction in territory of stenting artery, in-stent restenosis or occlusion was not demonstrated by emergency cerebral angiography. Conclusion: Interventional treatment of SISR is feasible and safe, however, further studies need to warrant the long-term effects.

Isoindoline nitroxide-labeled porphyrins as potential fluorescence-suppressed spin probes.

A series of isoindoline nitroxide-labeled porphyrins were synthesized by the reaction of 5-phenyldipyrromethane and 5-(4'-carboethoxy-methyleneoxyphenyl)dipyrromethane with 5-formyl-1,1,3,3-tetramethylisoindolin-2-yloxyl (FTMIO) using the Lindsey method. The corresponding water-soluble spin-labeled porphyrins were also prepared. Subsequently, these compounds were characterized and their in vitro properties were evaluated. The electrochemical assay demonstrated that these isoindoline nitroxide-labeled porphyrins had similar electrochemical and redox properties to 5-carboxy-1,1,3,3-tetramethylisoindolin-2-yloxyl (CTMIO). The electron paramagnetic resonance test showed that these porphyrins exhibited hyperfine splittings and characteristic spectra of CTMIO with typical nitroxide g-values and nitrogen isotropic hyperfine coupling constants. The in vitro cytotoxicity assay indicated that these porphyrins possessed low cytotoxicity to human renal tubular epithelial 293T cells (normal cells) and human hepatoma HepG2 cells (tumor cells). Fluorescence spectroscopy revealed that free base isoindoline nitroxide-labeled porphyrins exhibited fluorescence suppression characteristic of nitroxide-fluorophore systems. In vitro fluorescene imaging demonstrated that the reduced isoindoline nitroxide-labeled porphyrins eliminated fluorescence suppression and displayed strong red fluorescence imaging in HepG2 cells. Thus these isoindoline nitroxide-labeled porphyrins may be considered potentially as biological spin probes for fluorescence imaging and EPR spectroscopy.

Effect of Xianling Gubao capsules on bone mineral density in osteoporosis patients.

This paper aimed to analyze the clinical effects of Xianling Gubao capsules on the bone mineral density (BMD) of 94 patients with osteoporosis. After reviewing and analyzing the clinical information in our hospital from January 2015 to January 2016, the patients were divided into a control group and a treatment group with 47 cases in each group. Patients in the control group were treated with routine Western medicine, while the treatment group received Xianling Gubao capsules. Both groups were treated continuously for 3 courses (30 days each course) and had their BMD analyzed and compared. The effective rates of the treatment group and control group were 91.48% and 70.21%, respectively, with statistical significance (P less than 0.05). Compared with the same group before the treatment, the bone metabolic indexes (blood calcium, phosphorus, and alkaline phosphate) and the BMD of the femoral neck and lumbar vertebrae (L1-2, L3-4, L2-4) of both groups were all improved with statistical significance (P less than 0.05) after the treatment. The above indexes of the treatment group were all significantly higher than those of the control group, with statistical significance (P less than 0.05). Compared with the same group before treatment (P less than 0.05), the osteocalcin (OC) levels of both groups were increased, and the Cross-linked N-telopeptide of type 1 collagen (CTX-1) levels were decreased after the treatment. The OC level in the treatment group was higher when compared with the control group, while the CTX-1 level was lower compared to the control group (P less than 0.05). Moreover, no significant side-effects or adverse events were observed during the treatment and observation period. The Xianling Gubao Capsule possesses a therapeutic effect for BMD in osteoporosis patients, which can effectively increase their BMD, improve their bone metabolism, and control the loss of bone mass, therefore, can be used in clinical promotion and application.

Effects of Saccharomyces cerevisiae fermentation products on dairy calves: Performance and health.

The aim of the study was to evaluate the effects of Saccharomyces cerevisiae fermentation products (SCFP) on performance and health of calves during the first 63 d of age. Sixty Holstein calves (30 males and 30 females) at 2 d of age were blocked by sex and date of birth then randomly assigned within blocks to 1 of 3 treatments. A texturized calf starter was fed ad libitum containing 0 (control), 0.5, or 1% SCFP (Original XPC, Diamond V, Cedar Rapids, IA) of DM. In addition, the supplemented calves were fed 1 g/d SCFP (SmartCare, Diamond V) in milk until d 30. All calves were fed 4 L of colostrum within 1 h of birth and were subsequently fed milk twice daily until weaned at 56 d of age. Male calves were harvested on d 56. Performance and health of weaned female calves were monitored until 63 d of age to determine the effect of preweaning treatment of SCFP on weaning stress. Starter intake, fecal scores, and medical treatments were recorded daily. Body weight measures and blood samples were collected on d 2, 28, 56, and 63. Serum was analyzed for blood urea nitrogen, fatty acids, insulin-like growth factor-1, glucose, and total protein. Oxidative biomarkers and total antioxidant capacity were also evaluated in the serum. Body weight, DMI, blood parameters, and oxidative biomarkers did not differ among treatments. Supplementation of SCFP lowered fecal scores in the pre- and postweaning periods. Saccharomyces cerevisiae fermentation products can be used to reduce the diarrhea in calves grown under normal commercial conditions.