Subcutaneous lysophosphatidylcholine administration promotes a febrile and immune response in Holstein heifer calves.

Lysophosphatidylcholine (LPC) is immunomodulatory in nonruminants; however, the actions of LPC on immunity in cattle are undefined. Our objective was to study the effects of LPC administration on measures of immunity, liver health, and growth in calves. Healthy Holstein heifer calves (n = 46; age 7 ± 3 d) were randomly assigned to 1 of 4 treatments (n = 10 to 11 calves/treatment): a milk replacer diet unsupplemented with lecithin in the absence (CON) or presence of subcutaneously (s.c.) administered mixed (mLPC; 69% LPC-16:0, 25% LPC-18:0, 6% other) or pure LPC (pLPC; 99% LPC-18:0), or a milk replacer diet supplemented with 3% lecithin enriched in lysophospholipids containing LPC in the absence of s.c.-administered LPC (LYSO) for 5 wk. Calves received 5 s.c. injections of vehicle (10 mL of phosphate-buffered saline containing 20 mg of bovine serum albumin/mL; CON and LYSO) or vehicle containing mLPC or pLPC to provide 10 mg of total LPC per kilogram of BW per injection every 12 h during wk 2 of life. Calves were fed a milk replacer containing 27% crude protein and 24% fat at 1.75% of BW per day (dry matter basis) until wk 6 of life (start of weaning). Starter grain and water were provided ad libitum. Body measurements were recorded weekly, and clinical observations were recorded daily. Blood samples were collected weekly before morning feeding and at 0, 5, and 10 h, relative to the final s.c. injection of vehicle or LPC. Data were analyzed using a mixed model, with repeated measures including fixed effects of treatment, time, and their interaction. Dunnett's test was used to compare treatments to CON. Peak rectal temperatures were higher in mLPC or pLPC, relative to CON. Plasma LPC concentrations were greater in mLPC and LYSO calves 5 h and 10 h after the final injection, relative to CON. Calves receiving mLPC and pLPC also had higher circulating serum amyloid A concentrations, relative to CON. Calves receiving mLPC had greater serum aspartate aminotransferase, γ-glutamyltransferase, and glutamate dehydrogenase concentrations, relative to CON. Calves provided mLPC experienced lower average daily gain (ADG) after weaning, relative to CON. The LYSO treatment did not modify rectal temperatures, ADG, or measures of liver health, relative to CON. We conclude that LPC administered as s.c. injections induced an acute febrile response, modified measures of liver and immune function, and impaired growth in calves.

[Risk factors of cytomegalovirus infection in patients with failed corneal grafts].

Objective: To investigate the levels of cytomegalovirus (CMV) infection and associated risk factors in corneal transplant recipients who experienced transplant failure. Methods: This was a case-control study. Clinical data from 576 cases (576 eyes) of patients who underwent repeat corneal transplant surgery at the Department of Ophthalmology, Peking University Third Hospital, due to corneal transplant failure from January 2016 to May 2022 were collected. Of these, 305 were male and 271 were female, with a median age of 44.0 (0.7, 91.0) years. The CMV infection rate was analyzed based on the detection of CMV DNA in aqueous humor or corneal tissue during corneal transplant surgery. Patients were divided into the CMV group (CMV DNA positive) and the control group (herpes virus DNA negative). The main research indicators included the CMV infection rate, clinical characteristics, and risk factors in corneal transplant recipients. Chi-square tests and binary logistic analysis were used to compare differences between the two groups in general information, systemic diseases, ocular lesions, ocular surgical history, and local and systemic medications. Odds ratios (OR) and 95% confidence intervals (CI) were calculated for each CMV infection risk factor. Results: The overall CMV infection rate was 21.9%(126/576), with annual rates ranging from 10.9% to 37.7% from 2016 to 2021. After applying inclusion and exclusion criteria, 378 patients were included in the control trial, with 126 in the CMV group and 252 in the control group. Statistically significant differences between the two groups were observed in systemic immune-related corneal lesions [CMV group: 38 (30.2%), control group: 26 (10.3%)], local immune and inflammatory corneal lesions [CMV group: 46 (36.5%), control group: 40 (15.9%)], congenital corneal opacity [CMV group: 46 (36.5%), control group: 48 (19.0%)] total number of corneal transplants (CMV group: 178 times, control group: 276 times), corneal deep neovascularization crossing the graft [CMV group: 104 (82.5%), control group: 68 (27.0%)] and severe opacity [CMV group: 44 (34.9%), control group: 30 (11.0%)]. Binary logistic regression analysis showed that systemic immune-related corneal lesions (OR=4.044, 95%CI 1.810-9.033, P<0.001), local immune and inflammatory corneal lesions (OR=3.554, 95%CI 1.569-8.052, P=0.002), congenital corneal opacity (OR=2.606, 95%CI 1.216-5.589, P=0.014), total number of corneal transplants (OR=3.206, 95%CI 1.753-5.864, P<0.001), corneal deep neovascularization crossing the graft (OR=8.347, 95%CI 3.967-17.559, P<0.001), and severe opacity (OR=3.063, 95%CI 1.221-7.682, P=0.017) were independent risk factors for CMV infection after corneal transplant. Conclusions: CMV infection was present in more than 1/5 of corneal transplant recipients who experienced transplant failure. CMV infection after corneal transplant may be related to immune rejection reactions and ocular inflammatory responses. Inflammatory corneal lesions associated with systemic or local immune abnormalities, congenital corneal opacity, and multiple corneal transplants may exacerbate the levels of inflammatory factors during the perioperative period of corneal transplant, increasing the risk of post-transplant CMV infection, leading to the infiltration of deep neovascularization and severe opacity in the cornea.

Rheumatoid arthritis and the risk of major cardiometabolic diseases: a Mendelian randomization study.

OBJECTIVE: Rheumatoid arthritis (RA) is suggested to be implicated in the development of cardiometabolic diseases. We conducted a Mendelian randomization (MR) study to assess potential causality for associations of RA with the risk of cardiometabolic diseases, including type 2 diabetes (T2D), coronary artery disease (CAD), and ischaemic stroke. METHOD: Seventy independent single-nucleotide polymorphisms (SNPs) associated with RA were identified as instrumental variables from a genome-wide association study (GWAS) of 58 284 European subjects. Summary-level data for the associations of the 70 genetic variants with T2D, CAD, and ischaemic stroke were taken from three GWASs with a total of 1 529 131 participants. Inverse-variance weighted (IVW) MR was used in the main analyses. RESULTS: The main IVW MR analysis showed that genetically determined RA was associated with higher risks of T2D [odds ratio (OR): 1.04, 95% confidence interval (CI) 1.02-1.05; p < 0.001] and CAD (OR: 1.02, 95% CI 1.00-1.03; p = 0.012), but not ischaemic stroke (OR: 1.00, 95% CI 0.99-1.02; p = 0.961). Sensitivity analyses with multiple MR methods confirmed these associations. MR-Egger regression showed no evidence of pleiotropy in the association between genetically determined RA and the risk of T2D, CAD, and ischaemic stroke. Leave-one-out sensitivity analysis showed that the association between genetically determined RA and the risk of T2D, CAD, and ischaemic stroke was not driven by any individual SNP. CONCLUSION: Genetically determined RA was associated with increased risks of T2D and CAD, suggesting that RA plays a crucial role in the pathogenesis of T2D and CAD.

Replacing ground corn with soyhulls plus palmitic acid in low metabolizable protein diets with or without rumen-protected amino acids: Effects on production and nutrient utilization in dairy cows.

We previously observed that diets with reduced starch concentration decreased yields of milk and milk protein in dairy cows fed low metabolizable protein diets. Supplementation of reduced-starch diets with a lipid source may attenuate or eliminate production losses. Our objective was to investigate the effects of partially replacing ground corn with soyhulls plus a palmitic acid-enriched supplement on dry matter (DM) intake, milk yield and composition, plasma AA concentration, and N and energy utilization in cows fed low metabolizable protein diets (mean = -68 g/d balance) with or without rumen-protected Met, Lys, and His (RP-MLH). Sixteen multiparous Holstein cows averaging (mean ± standard deviation) 112 ± 28 d in milk, 724 ± 44 kg of body weight, and 46 ± 5 kg/d of milk in the beginning of the study were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d, consisting of 14 d for diet adaptation and 7 d for data and sample collection. Diets were fed as follows: (1) high starch (HS), (2) HS plus RP-MLH (HS+AA), (3) reduced starch plus a palmitic acid-enriched supplement (RSPA), and (4) RSPA plus RP-MLH (RSPA+AA). The HS diet contained (DM basis) 26% ground corn and 7% soyhulls, and the RSPA diet had 10% ground corn, 22% soyhulls, and 1.5% palmitic acid. The HS diet averaged (DM basis) 32.6% starch and 4% ether extract, while starch and ether extract concentrations of the RSPA diet were 21.7 and 5.9%, respectively. All 4 diets had (DM basis) 40% corn silage, 5% mixed-mostly grass haylage, 5% grass hay, and 50% concentrate. Diets did not affect DM intake and milk yield. Contrarily, feeding RSPA and RSPA+AA increased yields of energy-corrected milk (47.0 vs. 44.8 kg/d) and milk fat (1.65 vs. 1.50 kg/d) compared with HS and HS+AA. Milk fat concentration tended to decrease when RP-MLH was supplemented to HS, but no change was seen when added to RS (starch level × RP-MLH interaction). Milk and plasma urea N increased, and milk N efficiency decreased in cows fed RSPA and RSPA+AA versus HS and HS+AA. Apparent total-tract digestibilites of crude protein and neutral detergent fiber, as well as urinary urea N and total N excretion, were greater in cows offered RSPA and RSPA+AA than HS and HS+AA. Plasma Met and His concentrations increased with supplemental RP-MLH. Intake of gross energy and digestible energy and the output of urinary and milk energy were all greater with feeding RSPA and RSPA+AA versus HS and HS+AA. In summary, partially replacing ground corn with soyhulls plus palmitic acid in diets supplemented or not with RP-MLH increased milk fat yield and fiber digestibility and maintained DM intake and milk yield, but with decreased milk N efficiency and elevated urinary N excretion.

Comparison of anal function and quality of life after conformal sphincter preservation operation and intersphincteric resection of very low rectal cancer: a multicenter, retrospective, case-control analysis.

PURPOSE: Conformal sphincter preservation operation (CSPO) is a sphincter preservation operation for very low rectal cancers. Compared to intersphincteric resection (ISR), CSPO retains more dentate line and distal rectal wall, and also avoids damaging the nerves in the intersphincteric space. This study aimed to compare the postoperative anal function and quality of life between the CSPO and ISR. METHOD: Patients with low rectal cancer undergoing CSPO (n = 117) and ISR (n = 66) were included from Changhai and Huashan Hospital, respectively, between 2011 and 2020. A visual analog scale (range 0-10) was utilized to evaluate satisfaction with anal function and quality of life. The anal function was evaluated with Wexner scores and low anterior resection syndrome (LARS) score. Quality of life was evaluated with the EORTC QLQ-C30 and QLQ-CR38. RESULTS: The CSPO group had more male patients (65.8% vs. 50%, p = 0.042), more preoperative chemoradiotherapy (33.3% vs. 10.6%, p < 0.001), lower tumor position (3.45 ± 1.13 vs. 4.24 ± 0.86 cm, p < 0.001), and more postoperative chemotherapy (65% vs. 13.6%, p < 0.001) compared to the ISR group. In addition, CSPO patients had shorter postoperative stay (6.63 ± 2.53 vs. 7.85 ± 4.73 days, p = 0.003) and comparable stoma reversal rates within 1 year after surgery (92.16% vs. 96.97%, p = 0.318). Multivariable analysis showed that CSPO significantly contributed to higher satisfaction with anal function (beta = 1.752, 95% CI 0.776-2.728) and with quality of life (beta = 1.219, 95% CI 0.374-2.064), but not to Wexner, LARS score, or EORTC QLQ-C30 and QLQ-CR38. CONCLUSION: CSPO improved the satisfaction with anal function and quality of life but utilized more preoperative chemoradiotherapy. CSPO may be an alternative choice for patients with very low rectal cancers in better physical health and with higher requirements for anal function and quality of life.

Replacing soybean meal with okara meal: Effects on production, milk fatty acid and plasma amino acid profile, and nutrient utilization in dairy cows.

Okara meal is a byproduct from the production of soymilk and tofu and can potentially replace soybean meal (SBM) in dairy diets due to its high crude protein (CP) concentration and residual fat. The objective of this study was to investigate the effects of replacing SBM with okara meal on feed intake, yields of milk and milk components, milk fatty acid (FA) profile, nutrient utilization, and plasma AA concentration in lactating dairy cows. Twelve multiparous (65 ± 33 d in milk) and 8 primiparous (100 ± 35 d in milk) organically certified Jersey cows were paired by parity or days in milk, and within pair, randomly assigned to treatments in a crossover design with 21-d periods (14 d for diet adaptation and 7 d for data and sample collection). Diets were fed as total mixed ration formulated to be isonitrogenous and isofibrous and contained (dry matter basis) 50% mixed, mostly grass baleage, 2% sugarcane liquid molasses, 2% minerals-vitamins premix, and either (1) 8.1% SBM, 10% soyhulls, and 27.9% ground corn (CTRL); or (2) 15% okara meal, 8% soyhulls, and 23% ground corn (OKR). Dietary CP, ash-free neutral detergent fiber, and total FA averaged 15.4, 35.3, and 3.08% for CTRL and 15.9%, 36.3%, and 3.74% for OKR, respectively. Substitution of SBM with okara meal did not alter dry matter intake but increased intakes of CP and ash-free neutral detergent fiber. Additionally, no significant differences between treatments were observed for yields of milk and milk components, and concentrations of milk fat, lactose, and total solids. However, milk true protein concentration was lower in cows fed OKR (3.76%) versus CTRL (3.81%). Both milk urea N (8.51 vs. 9.47 mg/dL) and plasma urea N (16.9 vs. 17.8 mg/dL) concentrations decreased with OKR relative to the CTRL diet, respectively. Compared with CTRL, feeding OKR lowered the milk proportions of total odd-chain FA, de novo FA, and mixed FA and increased those of preformed FA, total n-6 FA, and total n-3 FA. The milk proportions of trans-10 18:1, trans-11 18:1, and cis-9,trans-11 18:2 were greater with feeding OKR versus the CTRL diet. The apparent total-tract digestibility of nutrients, urinary excretion of total purine derivatives (uric acid plus allantoin), and total N were not affected by treatments. Except for plasma Leu, which was lower in OKR compared with the CTRL diet, no other significant changes in the plasma concentrations of AA were observed. The plasma concentration of carnosine was lowest in cows receiving the OKR diet. Overall, our results revealed that okara meal can completely replace SBM without negatively affecting production and nutrient digestibility in early- to mid-lactation Jersey cows. Further research is needed to assess the economic feasibility of including okara meal in dairy diets, as well as the amount of okara meal that maximizes yields of milk and milk components in dairy cows in different stages of lactation.

Dietary starch level and rumen-protected methionine, lysine, and histidine: Effects on milk yield, nitrogen, and energy utilization in dairy cows fed diets low in metabolizable protein.

Our objective was to investigate the interactions between starch level and rumen-protected Met, Lys, His (RP-MLH) on milk yield, plasma AA concentration, and nutrient utilization in dairy cows fed low metabolizable protein diets (mean = -119 g/d of metabolizable protein balance). Sixteen multiparous Holstein cows (138 ± 46 d in milk, 46 ± 6 kg/d in milk) were used in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments. Each period lasted 21 d with 14 d for diet adaptation and 7 d for data and sample collection. Dietary starch level varied by replacing (dry matter basis) pelleted beet pulp and soyhulls with ground corn resulting in the following treatments: (1) 20% pelleted beet pulp and 10% soyhulls (reduced starch = RS), (2) RS plus RP-MLH (RS+AA), (3) 30% ground corn (high starch = HS), and (4) HS plus RP-MLH (HS+AA). Dietary starch concentrations averaged 12.3 and 34.4% for RS and HS basal diets, respectively. Diets were supplemented with RP-MLH products to supply digestible Met, Lys, and His. Compared with RS and RS+AA diets, HS and HS+AA diets increased yields of milk (37.9 vs. 40.1 kg/d) and milk protein (1.07 vs. 1.16 kg/d) and decreased dry matter intake (25.9 vs. 25.2 kg/d), milk urea N (12.6 vs. 11.0 mg/dL), and plasma urea N (13.3 vs. 11.6 mg/dL). Milk N efficiency was greater in cows fed the HS and HS+AA than RS and RS+AA diets (28.9 vs. 25%), and RP-MLH supplementation improved milk true protein concentration. Starch level × RP-MLH interactions were observed for plasma concentrations of Arg and Lys, with RP-MLH being more effective to increase plasma Arg (+16%) and Lys (+23%) when supplemented to the RS than the HS basal diet. Replacing pelleted beet pulp and soyhulls with ground corn lowered the plasma concentrations of all essential AA except Met and Thr. In addition, the plasma concentrations of His and Met increased with RP-MLH. The apparent total-tract digestibilities of neutral and acid detergent fiber were lower, and those of starch and ether extract greater in cows offered the HS and HS+AA diets than RS and RS+AA diets. Urinary excretion of urea N decreased by replacing pelleted beet pulp and soyhulls with ground corn. Enteric CH4 production, CH4 yield, and CH4 intensity all decreased in the HS and HS+AA versus RS and RS+AA diets. Diets did not affect the intakes of gross energy, metabolizable energy, and net energy of lactation. In contrast, digestible energy intake increased with feeding the RS and RS+AA diets, whereas CH4 energy decreased in cows fed the HS and HS+AA diets. Supplementation with RP-MLH had no effect on energy utilization variables. Overall, the lack of interactions between dietary starch level and RP-MLH supplementation on most variables measured herein showed that the effects of starch intake and RP-MLH were independent or additive.

[The expression of GGCT in the bladder urothelial cell carcinoma and its clinical significance].

Objective: To evaluate the expression and clinical significance of γ-glutamylcyclotransferase (GGCT) in patients with bladder urothelial cell carcinoma. Methods: Immunohistochemical staining for GGCT were performed on tissue sections of 86 patients with bladder urothelial cell carcinoma and 10 normal controls, and the correlations between GGCT and clinicopathological characteristics and the prognosis were analyzed. Results: The positive rate of the expression of GGCT in 86 cases of bladder urothelial cell carcinoma was 61.6% (53/86). GGCT protein was located mainly in cancer cell cytoplasm, and it can be seen in the nucleus of the tumor cells in some cases. The level of GGCT expression was positively related to pathological classification (P<0.001), stage (P=0.020), and tumor size (P=0.025). Immunohistochemical semiquantitative analysis showed that the expression of GGCT in patients with T1 stage of non-muscle invasion bladder urothelial cell carcinoma was significantly higher than that with Ta stage (P=0.034). Kaplan-Meier analysis showed that the expression of GGCT was correlated with the recurrence-free survival in patients with non-muscle invasive bladder cancer, the recurrence-free survival rate was lower in the GGCT positive group (P=0.029). Multivariate COX regression analysis showed that the pathological stage (OR=5.029, P=0.009) and the number of tumors (OR=3.320, P=0.024)were the independent risk factors for recurrence-free survival in patients with early urothelial cell carcinoma of the bladder. Conclusions: The expression of GGCT is significantly increased in bladder urothelial cell carcinoma and is related to the malignant biological behavior and progression of tumor. Patients with GGCT positive early bladder tumor are inclined to recur.

[Chlorpromazine hydrochloride plays a tumor suppressive role in diffuse large B lymphoma by promoting the expression of S1PR2].

Objective: To investigate the effect of chlorpromazine hydrochloride on proliferation, apoptosis and cell cycle arrest of human diffuse large B lymphoma cells and its possible mechanism. Methods: In January 2019, OCI-LY3 and TMD8 cells were treated with different concentrations of chlorpromazine hydrochloride for 72 h, the proliferations were detected by Alamar Blue assay. The apoptosis and cell cycle arrest of OCI-LY3 and TMD8 cells were detected by flow cytometry with FITC annexin V/PI (propidium iodide) double staining and PI, respectively. The mRNA levels of cyclin-dependent kinase inhibitor P21, P27, CyclinD1 and S1PR2 were detected by RTqPCR. The protein levels of P21, P27 and S1PR2 were detected by Western blot. Results: With the increase of chlorpromazine hydrochloride concentration, the proliferation inhibition rates of OCI-LY3 and TMD8 cells increased at 72 hours, apoptosis and G1 cell cycle arrest increased at 24 hours. Compared with the control group, the expression levels of P21, P27 and SIPR2 mRNA in OCI-LY3 and TMD8 cells were increased at 10 µmol/L chlorpromazine after 12 hours treatment (P<0.05) . And there was no significant difference in the expression of CyclinD1 mRNA (P>0.05) . There was a similar increase in protein levels of P21, P27 and SIPR2 in OCI-LY3 and TMD8 cells after 24 hours of treatment (P<0.05) . Conclusion: Chlorpromazine hydrochloride at a specific concentration may inhibit the proliferation of ABC diffuse large B lymphoma cells by promoting the expression of S1PR2, and promote cell apoptosis and G1 phase cell cycle arrest.

A systematic review of clinical practice guidelines on uncomplicated birth.

BACKGROUND: Clinical practice guidelines on uncomplicated birth provide clinical practice guidance and help to reduce unnecessary, non-evidence-based and potentially harmful intrapartum care practices. Little is known about the trustworthiness and consistency of these guidelines. OBJECTIVES: To appraise guidelines relevant to uncomplicated birth, and summarise consensus and non-consensus recommendations. SEARCH STRATEGY: Eight literature databases and the websites of guideline development institutions and organisations of obstetricians, gynaecologists and midwives were searched from January 2008 to October 2018. SELECTION CRITERIA: Guidelines that: reported in Chinese or English; labelled guideline, or recommendation, or consensus, or practice parameter, or position paper/stand; with uncomplicated birth being the primary objectives or comprised chapter(s); and were the most recently published or updated versions. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed guideline quality using the AGREE II instrument, and synthesised consensus and non-consensus recommendations using the content analysis approach. MAIN RESULTS: Eleven guidelines met the inclusion criteria. The WHO and NICE guidelines were deemed to have the highest methodological quality. Twenty-three discrepant recommendations and 39 groups of unanimous recommendations (containing 113 individual ones) were identified, among which 14 recommendations including eating and drinking as desired, prophylactic uterotonics, routine amniotomy and episiotomy were included in five or more guidelines. Perineal massage, uterine massage, active or expectant management at the third stage and use of hands-on or hands-poised technique were identified as the main discrepant recommendations. CONCLUSIONS: Two guidelines with higher methodological quality and key clinical guideline recommendations, including both consensus and non-consensus ones, on uncomplicated birth were identified. TWEETABLE ABSTRACT: Clinical guidelines of uncomplicated birth agree and disagree on several key recommendations.

Study on the characteristics and mechanism of pulsed laser cleaning of polyacrylate resin coating on aluminum alloy substrates.

The behavior of paint removal with a pulsed laser has been investigated using an Nd:YAG fiber laser. Experimental and theoretical analyses are conducted to reveal the underlying mechanism of the paint removal. The results show that the depth, radius, and volume of the pit formed by a single pulse become larger with increases in the energy density. The ideal parameters for the complete removal of paint have been achieved. During the paint removal, burning occurred over the surface, and the variation in element content has been attributed to the absorption of laser energy. Under the action of the pulsed laser, there was breakage and rearrangement of chemical bonds such as C-C, C-N, and C-O in the molecular chain of the polyacrylate paint coating. Through these analyses, the paint removal mechanism was shown to be based on chemical bond breakage, combustion, and mechanical action provided by the thermal expansion and plasma shock.

The acute phase protein orosomucoid 1 is upregulated in early lactation but does not trigger appetite-suppressing STAT3 signaling via the leptin receptor.

Dairy cows consume inadequate amounts of feed in early lactation and during conditions and diseases such as excessive fatness, heat stress, and infectious diseases. Affected cows often experience increases in plasma concentrations of acute phase proteins consistent with the negative effect of inflammation on appetite. The acute phase protein orosomucoid 1 (ORM1), also known as alpha-1-acid glycoprotein, was recently reported to reduce appetite in the mouse through its ability to bind the full-length leptin receptor (Ob-Rb) and activate appetite-suppressing signal transducer and activator of transcription 3 (STAT3) signaling. These observations raise the possibility that ORM1 exerts appetite-suppressing effects in dairy cattle during periods of increased inflammatory tone. The applicability of this model was assessed in 2 ways. First, we asked whether ORM1 is regulated during periods of inadequate appetite such as the transition from late pregnancy to early lactation and periods of increased inflammatory tone. Plasma ORM1 was invariant in late pregnancy but increased 2.5-fold between parturition and d 7 of lactation. Gene expression studies showed that liver was the major source of this elevation with little contribution by adipose tissue or mammary gland. Additional studies showed that plasma ORM1 was not increased further by excessive fatness or by reproductive dysfunction in early lactation and was completely unresponsive to inflammatory stimuli such as heat stress or intravascular administration of the endotoxin lipopolysaccharide during established lactation. Second, we tested the ability of ORM1 to trigger STAT3 signaling through Ob-Rb using Chinese hamster ovary K1 (CHO-K1) cells transfected with a STAT3 expression plasmid. In this configuration, CHO-K1 cells did not express Ob-Rb and were incapable of leptin-induced STAT3 phosphorylation. Leptin responsiveness was conferred by co-transfecting with bovine Ob-Rb, with leptin causing increases of 5.7-fold in STAT3 phosphorylation and 2.1-fold in the expression of the STAT3-dependent gene, SOCS3. In contrast, neither bovine or human ORM1 triggered STAT3 phosphorylation irrespective of dose and period of incubation tested. In summary, bovine ORM1 is not increased during periods of increased inflammatory tone except in early lactation and is incapable of Ob-Rb-dependent STAT3 signaling. Overall, these data are inconsistent with ORM1 mediating the appetite-suppressing effects of inflammation in cattle through Ob-Rb.

MicroRNA-101 regulates autophagy, proliferation and apoptosis via targeting EZH2 in laryngeal squamous cell carcinoma.

Laryngeal carcinoma is the second commonest head and neck carcinoma globally. MicroRNA-101 (miR-101) has been reported as a tumor suppressor in multiple malignancies including laryngeal squamous cell carcinoma (LSCC). However, the roles and molecular mechanisms of miR-101 in the development of LSCC have not been fully elucidated. In present study, RT-qPCR assay was performed to detect the expression of miR-101 and enhancer of zeste homolog 2 (EZH2) mRNA. Western blot assay was conducted to determine protein expression of LC3-Ⅰ, LC3-Ⅱ, p62 and EZH2. Cell proliferative capacity was evaluated by MTS assay. The effect of miR-101 alone or along with EZH2 on cell apoptosis was assessed by apoptotic index and caspase-3 activity. Bioinformatic analysis, luciferase assay and RNA immunoprecipitation (RIP) assay were carried out to investigate the interaction between miR-101 and EZH2. Results revealed that miR-101 expression was strikingly down-regulated in LSCC cell lines. Functional analyses showed that ectopic expression of miR-101 suppressed cell autophagy and proliferation and facilitated cell apoptosis in LSCC. Further investigations revealed that miR-101 inhibited EZH2 expression by direct interaction and EZH2 was highly expressed in LSCC cells. Also, EZH2 knockdown reduced the autophagic activity of LSCC cells. Moreover, restoration experiments showed that EZH2 up-regulation weakened miR-101-mediated anti-autophagy, anti-proliferation and pro-apoptosis effects in LSCC cells. In conclusion, our findings suggested that miR-101 inhibited autophagy and proliferation and promoted apoptosis via targeting EZH2 in LSCC, providing a deep insight into the pathogenesis of LSCC and hinting the pivotal roles of epigenetic modifications especially histone methylation in the development of LSCC.

Incremental amounts of rumen-protected histidine increase plasma and muscle histidine concentrations and milk protein yield in dairy cows fed a metabolizable protein-deficient diet.

The dairy industry can benefit from low crude protein (CP) diets due to reduced N excretion, but shortages of Met, Lys, and His may limit milk protein synthesis. We studied the effect of incremental amounts of rumen-protected (RP)-His on plasma and muscle AA profile, nutrient utilization, and yields of milk and milk true protein in dairy cows. Eight multiparous Holstein cows (130 ± 30 d in milk) were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with 28-d experimental periods. Treatments included a basal diet composed (dry matter basis) of 50% corn silage, 15% haylage, and 35% concentrate supplemented with 0, 82, 164, and 246 g/d of RP-His and 11 g/d of RP-Met. Milk, plasma, and muscle samples were collected weekly or every other week during all 4 periods, whereas spot urine and fecal grab samples were taken only in wk 4 of each period. Data were analyzed individually by week using linear, quadratic, and cubic orthogonal polynomials and repeated measures. Plasma His increased linearly with RP-His during wk 1 (30.3 to 57.2 µM) to wk 4 (33.2 to 63.1 µM). Plasma carnosine increased linearly with supplemental RP-His except in wk 2. No treatment effect was observed for plasma 3-methylhistidine except a quadratic effect in wk 3. Inclusion of RP-His showed linear effects on muscle His in wk 2 (20.1 to 32.5 µM) and 4 (20.3 to 35.5 µM). Whereas muscle anserine and carnosine concentrations were not affected by treatments in wk 4, anserine responded quadratically and carnosine showed a trend for a quadratic response to RP-His in wk 2. During wk 4, treatments did not affect urinary excretion of total purine derivatives, as well as dry matter intake and milk concentrations of fat and true protein. In contrast, milk yield tended to increase linearly (31.2 to 32.7 kg/d) and milk true protein yield responded linearly (0.93 to 0.98 kg/d) and tended to increase quadratically to RP-His supplementation in wk 4. Also, milk urea-N (11.7 to 12.9 mg/dL) and urinary excretion of urea-N (23.7 to 27.0% of N intake) increased linearly with feeding RP-His in wk 4. Overall, RP-His was effective to enhance plasma and muscle concentrations of His and milk protein synthesis. Elevated milk urea-N and urinary excretion of urea-N suggest that plasma His may have exceeded the requirement with excess N converted to urea in the liver. Future research is needed to determine the bioavailability of RP-His supplements to improve the accuracy of diet formulation for AA.

Methyl donor supplementation suppresses the progression of liver lipid accumulation while modifying the plasma triacylglycerol lipidome in periparturient Holstein dairy cows.

Co-supplementation of methyl donors may lower hepatic lipid content in transition cows. To define the ability of methyl donor supplementation (MDS) to reduce hepatic lipid content and modify the plasma lipidome, 30 multiparous Holstein cows (2.04 ± 0.69 lactations; 689 ± 58 kg of body weight; 3.48 ± 0.10 units of body condition score) were fed a ration with or without rumen-protected methyl donors (22 g/d of Met, 10 g/d of choline chloride, 3 g/d of betaine, 96 mg/d of riboflavin, and 1.4 mg/d of vitamin B12) from d -28 before expected calving through d 14 postpartum. Cows were randomly enrolled based on predefined selection criteria (body condition score and parity). Base diets without MDS were formulated for gestation (15.4% crude protein with a predicted Lys-to-Met ratio of 3.25; 1.44 Mcal of net energy for lactation/kg of dry matter) and lactation (16.6% crude protein with a predicted Lys-to-Met ratio of 3.36; 1.64 Mcal of net energy for lactation/kg of dry matter). Blood sampling occurred from d -28 relative to expected calving through d 14 postpartum. Liver tissue was biopsied at d -28 relative to expected calving and on d 5 and 14 postpartum. In addition to routine analyses, serum AA concentrations on d 10 and 12 were quantified using mass spectrometry. Plasma triacylglycerol (TAG) and cholesteryl esters (CE) were qualitatively measured using time-of-flight mass spectrometry. Data were analyzed using a mixed model with repeated measures. Dry matter intake and milk yield were not modified by MDS. The transition from d -28 relative to expected parturition to d 14 postpartum was characterized by increased plasma fatty acid (0.15 to 0.71 mmol/L) and ß-hydroxybutyrate (0.34 to 0.43 mmol/L) levels and liver lipid content (3.91 to 9.16%). Methyl donor supplementation increased the serum Met level by 26% and decreased the serum Lys-to-Met ratio by 21% on d 10 and 12, respectively. Moreover, the increase in hepatic lipid content from d 5 through 14 postpartum was suppressed with MDS relative to control (3.57 vs. -0.29%). Dietary MDS modified the TAG and CE lipidome. For example, MDS increased plasma TAG 46:3 (carbon number:double bond) by 116% relative to control cows on d 5 postpartum. Moreover, MDS tended to increase plasma CE 34:6. In contrast, MDS lowered plasma TAG 54:8 by 39% relative to control cows on d 5 postpartum. We concluded that in the absence of gains in dry matter intake and milk and milk protein yields, dietary MDS slows the progression of hepatic lipid accumulation and modifies the plasma TAG lipidome in transition cows.

TLR5 activation induces expression of the pro-inflammatory mediator Urokinase Plasminogen Activator via NF-κB and MAPK signalling pathways in human dental pulp cells.

AIM: To explore the involvement of TLR5 in pulp inflammation and to examine the effects of TLR5 activation with its ligand, FlaB protein, on pro-inflammatory gene expression. METHODOLOGY: TLR5 expression in dental pulp tissues and human dental pulp cells (hDPCs) were determined by immunohistochemistry, immunocytochemistry, Western blots and RT-PCR analyses. To examine the role of TLR5, hDPCs were treated with recombinant FlaB protein (500 ng mL-1 ) to activate the receptor or with a small interfering RNA against TLR5 (si-TLR5) to downregulate the receptor. After exposure to FlaB, the expression of inflammation-related proteins was screened using a protein array kit. Western blots or qRT-PCR analyses were performed to identify changes in the expression of uPA (urokinase plasminogen activator), TIMPs (tissue inhibitor of metalloproteinases), and IL-6 and to determine their signalling pathways. Statistical analysis was performed using one-way analysis of variance (anova) with Tukey post hoc test; P < 0.05 was considered statistically significant. RESULT: TLR5 expression was identified in pulp tissues and hDPCs. In the protein array analysis, treatment with FlaB significantly increased uPA expression (P < 0.01) and significantly decreased TIMP1/4 (P < 0.05). FlaB treatment also significantly increased expression of the inflammatory marker IL-6 (P < 0.01). FlaB treatment increased phosphorylation of the NF-κB p65 subunit, JNK, p38 and ERK. Chemical inhibitors of NF-κB (Bay11-7082), p38 (SB202190) or ERK (U0126) decreased the FlaB induction of uPA expression. Downregulation of TLR5 expression by siRNA decreased the FlaB induction of uPA protein and p65 phosphorylation. CONCLUSION: TLR5 activation with FlaB treatment induced the expression of uPA via the NF-κB and MAPK signalling pathways. Flagellin-bearing oral bacteria may cause pulp inflammation through TLR5. The findings provide new clues to control pulpal diseases by targeting TLR5 signalling pathways.

Influence of olfactory dysfunction on the perception of food.

PURPOSE: Eating-related problems are among the most frequent issues in olfactory impairment, causing a noticeable loss of quality of life for some of the affected persons. To what extent olfactory dysfunction impacts on the sensory perception of food is less explored. The aim of the present study was to examine the impact of olfactory dysfunction on the perception of food aromas, as well as the perception of the "basic tastes" salty, sour, sweet, and bitter. METHODS: Eighty-nine participants were recruited for the prospective study. Group 1 consisted of thoroughly examined patients with olfactory dysfunction (n = 48, mean age = 60.0 years), group 2 consisted of people with normal olfactory function (n = 41, mean age = 50.4 years). First, olfactory and gustatory functions were assessed for all participants with the help of the "Sniffin'Sticks" battery and the "taste strips" test. Second, food odors were rated for their pleasantness, intensity, familiarity and desirability. Last, real food items were tasted orally and the intensity for basic taste qualities (sweet, bitter, salty, and sour) and pleasantness was rated. In addition, salivation was measured following exposure to the food odors. RESULTS: In comparison to controls, patients rated orthonasal food odors as less pleasant, intense, familiar, and less appetizing. "Taste strip" scores were significantly lower in patients (M = 9.56, SD = 2.76) as compared to controls (M = 10.88, SD = 1.89). In addition, ratings of food liking for chocolate and peanut were lower in patients compared to controls (chocolate: patients-M = 6.85, SD = 2.09, controls-M = 7.90, SD = 1.53; peanut: patients-M = 4.88, SD = 2.20, controls-M = 6.80, SD = 2.33). No significant differences were found regarding the comparison of the salivary flow rate in controls (M = 0.52 g/min, SD = 0.19) and patients (M = 0.50 SD = 0.17). CONCLUSIONS: Changes in the perception of odors may change the perception of food with specific effects on food liking. Olfactory dysfunction affects gustatory function, indicating the central-nervous interaction between taste and smell. Still, olfactory dysfunction did not appear to affect patients' salivary flow.

[Study on the relationship between preoperative evaluation, operation and prognosis of microvascular decompression for the treatment of hemifacial spasm].

Objective: To investigate the relationship between preoperative evaluation, surgery and prognosis of microvascular decompression (MVD) for the treatment of hemifacial spasm (HFS). Methods: The clinical data of 128 HFS patients treated with MVD in the department of neurosurgery of Taizhou Hospital of Zhejiang Province were retrospectively analyzed. According to the SMC grading system, the patients were divided into general spasm group and severe spasm group, and the clinical characteristics, offending vessel, prognosis and surgical complications of the two groups were compared. Results: In the general spasm group,the age at MVD was (48.6±10.6) years, the disease duration was (4.2±3.3) years;while in the severe spasm group,the age at MVD was (51.8±9.9) years, the disease duration was (8.1±4.5) years;the differences of age and disease duration between the two groups were statistically significant (P<0.05).In the general spasm group, there were 41 cases in which the offending vessel were AICA, 21 cases were PICA, 1 case was VA, 63 cases were single offending vessel, and 7 cases were multiple offending vessels. In the severe spasm group, there were 29 cases in which the offending vessel were AICA, 13 cases were PICA, 2 cases were VA, the total of 44 cases were single offending vessel and 14 cases were multiple offending vessels.There was a significant difference in the proportion of multiple offending vessels in the two groups, and the difference was statistically significant (P<0.05).Patients in the two groups were followed up for 12 to 32 months after surgery, and the difference in effective rate and recurrence rate was not statistically significant (P>0.05).Some kinds of postoperative complications were different between the two groups, the incidence of postoperative delayed facial paralysis was statistically significant (P<0.05), and the other complications were not statistically significant (P>0.05). Conclusion: Compared with the general spasm group, the patients in the severe spasm group were older, with longer disease duration, higher probability of multiple offending vessels and higher incidence of postoperative delayed facial paralysis. Therefore, preoperative SMC grading is helpful for the evaluation and prediction of intraoperative and postoperative conditions, which is worthy of wide clinical application.

[Clinical efficacy of different vascular reconstruction on adult moyamoya disease].

Objective: To investigate the clinical efficacy of superficial temporal artery -middle cerebral artery combined with encephalo-duro-arterio-myo-synangiosis (STA-MCA+EDAMS) and encephalo-duro-arterio-myo-synangiosis (EDAMS) in the treatment of adult moyamoya disease. Methods: The clinical data of 47 adult patients with moyamoya disease who received vascular reconstruction in the Department of Neurosurgery of Taizhou Hospital of Zhejiang Province from January 2014 to January 2018 were retrospectively analyzed. Among them, 21 patients received EDAMS alone (EDAMS group, 14 patients with hemorrhagic moyamoya disease, 7 patients with ischemic moyamoya disease), 26 patients received STA-MCA combined with EDAMS (STA-MCA+EDAMS group, 17 patients with hemorrhagic moyamoya disease, 9 patients with ischemic moyamoya disease). Cerebral hemodynamics at 1 day before surgery and 3 and 6 months after surgery were compared. The clinical efficacy and postoperative complications of the two methods were compared at 3 and 6 months postoperatively in hemorrhagic and ischemic types. Results: For hemorrhagic moyamoya disease, the remission rate (94.1%) at 6 months after surgery in the STA-MCA + EDAMS group was higher than that in the EDAMS group (57.1%), and the difference was statistically significant (P<0.05). The CBF and CBV in the STA-MCA+EDAMS group were higher than those in the EDAMS group at 3 and 6 months after operation, and the MTT and TPP were lower than those in the EDAMS group, but there was no significant difference between the two groups (all P>0.05). For hemorrhagic moyamoya disease and ischemic moyamoya disease, the total incidence of postoperative complications of the two surgical methods was different, but the difference was not statistically significant (both P>0.05). Conclusion: Superficial temporal artery -middle cerebral artery combined with encephalo-duro- arterio-myo-synangiosis (STA-MCA+EDAMS) and encephalo-duro-arterio-myo-synangiosis (EDAMS) can significantly improve neurological function and cerebral hemodynamics in adult moyamoya disease patients with high safety.

Short communication: Circulating fatty acylcarnitines are elevated in overweight periparturient dairy cows in association with sphingolipid biomarkers of insulin resistance.

Defects in mitochondrial fatty acid processing are associated with the development of fatty liver disease, inflammation, and insulin resistance in overweight nonruminants. Surplus fatty acids (FA) and defects in FA oxidation favor the accumulation of fatty acylcarnitines (FAC) and the sphingolipid ceramide. Moreover, elevated circulating FAC and ceramide concentrations are inversely related to insulin action. Because we have previously demonstrated that plasma ceramide levels increase during the transition from gestation to lactation, our aim was to determine whether changes in plasma medium- and long-chain FAC levels are related to circulating FA and sphingolipids in peripartal dairy cows. We hypothesized that plasma FAC levels would be higher in overweight cows experiencing increased lipolysis, and that FAC levels would be positively associated with elevations in plasma ceramides. Twenty-one multiparous Holstein cows were grouped according to body condition score (BCS) at d -30 prepartum as lean (BCS <3.0; n = 10) or overweight (BCS >4.0; n = 11). Blood was collected at d -30, -15, -7, and 4, relative to parturition. Circulating FAC and ceramide levels were determined using liquid chromatography and tandem mass spectrometry. To investigate the potential contributions of sphingomyelin (SM) hydrolysis to ceramide accrual, we also determined plasma SM levels during the peripartum period. Data were analyzed under a mixed model with the fixed effects of adiposity and time, and the random effect of cow. Relative to lean cows, overweight cows had elevated FAC during the transition from gestation to lactation. Circulating FAC levels were positively associated with FA, ceramide, and dihydro-SM levels. Although circulating FAC levels increased in all cows during the peripartum, enhanced prepartum adiposity contributed to a greater rise in plasma FA and FAC. Our results support on-going efforts to determine whether altered mitochondrial FA processing promotes the accumulation of the insulin resistance biomarker ceramide in blood and liver.