Pharmacological Mechanisms of Kirenol against Ovarian Carcinoma: A Network Pharmacology and Experimental Validation Study In Vitro.

BACKGROUND: Ovarian carcinoma is an aggressive gynecological malignancy. Kirenol, a diterpene compound, has recently gained attention for its potential anticancer properties. However, its exact anti-tumor mechanism remains largely unexplored. OBJECTIVE: In this study, we explored the inhibitory effects of Kirenol on ovarian cancer using network pharmacology and in vitro experiments and elucidated its underlying mechanisms. METHODS: Through the utilization of molecular docking, we established a network of proteinprotein interactions (PPI), which unveiled CDK4 as an essential target. Additionally, gene enrichment and pathway analysis highlighted the significance of the PI3K/AKT pathway. The viability of ovarian cancer cells and normal ovarian epithelial cells was evaluated using CCK8 assays to determine the effect of Kirenol. Following in vitro tests, cell colony formation, wound healing, flow cytometry, and Western blotting were conducted to assess its impact on cell proliferation, metastasis, apoptosis, and the cell cycle. RESULTS: Kirenol significantly reduced the viability of ovarian cancer cells (SKOV3 and A2780) compared to normal ovarian epithelial cells (IOSE-80). Moreover, Kirenol efficiently suppressed the growth and movement, caused a cell cycle halt, and stimulated programmed cell death in SKOV3 and A2780 cells. Through molecular analysis, it was observed that Kirenol increased the expression of Bax while decreasing the expression of MMP2, MMP9, and Bcl-2. It also attenuated the phosphorylation of PI3K, AKT, and RB and downregulated CDK4 and CCND1 expression. Notably, co-treatment with the PI3K pathway inhibitor LY294002 enhanced the inhibitory effect of Kirenol on ovarian cancer cells. CONCLUSION: In summary, the combined results of our network pharmacology analysis and in vitro tests emphasized that Kirenol hinders the growth of ovarian cancer cells, causes cell cycle arrest, enhances apoptosis, and hampers migration, possibly by regulating the PI3K/AKT/CDK4 signaling pathway.

Mechanisms of autophagy and endoplasmic reticulum stress in the reversal of platinum resistance of epithelial ovarian cancer cells by naringin.

OBJECTIVE: Our previous studies showed that naringin (Nar) can effectively reverse the cisplatin resistance of ovarian cancer cells. This study aims to explore the potential mechanism by which Nar reverses cisplatin resistance in ovarian cancer. METHODS: The proliferative activity of cells was evaluated using CCK8 and cell clone formation assays. Autophagic flux in cells was evaluated via LC3B immunofluorescence and monodansylcadaverine (MDC) staining. The expression levels of autophagy, endoplasmic reticulum (ER) stress, and apoptosis-related proteins were detected via Western blotting. Autophagy and ER stress were regulated using siATG5, siLC3B, rapamycin (Rap), chloroquine (CQ), 4-phenylbutyric acid (4-PBA), and thapsigargin (TG). siATG5 and siLC3B are short interfering RNAs (siRNAs) used to knock down the expression of ATG5 and LC3B genes, respectively. RESULTS: Nar inhibited autophagy in SKOV3/DDP cells by activating the PI3K/AKT/mTOR pathway. And Nar increased the levels of ER stress-related proteins, namely, P-PERK, GRP78, and CHOP, and promoted apoptosis in SKOV3/DDP cells. Moreover, treatment with the inhibitor of ER stress alleviated apoptosis induced by Nar in SKOV3/DDP cells. In addition, compared to cisplatin or naringin alone, the combination of Nar and cisplatin significantly reduced the proliferative activity of SKOV3/DDP cells. And siATG5, siLC3B, CQ or TG pretreatment further inhibited the proliferative activity of SKOV3/DDP cells. Conversely, Rap or 4-PBA pretreatment alleviated the cell proliferation inhibition caused by Nar combined with cisplatin. CONCLUSION: Nar not only inhibited the autophagy in SKOV3/DDP cells by regulating the PI3K/AKT/mTOR signalling pathway, but also promoted apoptosis in SKOV3/DDP cells by targeting ER stress. Nar can reverse the cisplatin resistance in SKOV3/DDP cells through these two mechanisms.

Recombinant Expression of Human IL-33 Protein and Its Effect on Skin Wound Healing in Diabetic Mice.

Chronic refractory wounds are one of the complications of diabetes mellitus that require effective therapy. The dermal-wound-healing property of IL-33 in diabetics is little understood. Therefore, this study aimed to express recombinant humanized mature IL-33 (rhmatIL-33) in Escherichia coli BL21 (DE3) and demonstrate its efficacy on dermal wounds in streptozotocin (STZ)-induced diabetic and nondiabetic mice by the dorsal incisional skin wound model. Results revealed that the rhmatIL-33 accelerated the scratch-healing of keratinocytes and fibroblasts at the cellular level. The wounds of diabetic mice (DM) showed more severe ulceration and inflammation than wild-type mice (WT), and the exogenous administration of rhmatIL-33 increased wound healing in both diabetic and wild-type mice. Compared with the up-regulation of endogenous IL-33 mRNA after injury in WT mice, the IL-33 mRNA decreased after injury in DM mice. Exogenous IL-33 administration increased the endogenous IL-33 mRNA in the DM group but decreased the IL-33 mRNA expression level of the WT group, indicating that IL-33 plays a balancing role in wound healing. IL-33 administration also elevated ILC2 cells in the wounds of diabetic and non-diabetic mice and improve the transcript levels of YM1, a marker of M2 macrophages. In conclusion, Hyperglycemia in diabetic mice inhibited the expression of IL-33 in the dermal wound. Exogenous addition of recombinant IL-33 promoted wound healing in diabetic mice by effectively increasing the level of IL-33 in wound tissue, increasing ILC2 cells, and accelerating the transformation of macrophage M1 to M2 phenotype.

Effect of Low-Frequency Electrical Stimulation Combined with Tonifying Kidney and Blood Pills on Uterine Rejuvenation after Abortion.

Objective: This study aimed to observe the clinical efficacy of low-frequency electrical stimulation therapy combined with tonifying the kidney and activating blood pills to promote uterine recovery after abortion and its effect on heat-shock protein (HSP)70 and HSP90. Methods: All cases were women with early pregnancy who underwent an abortion at the Third Affiliated Hospital of Nanchang University from September 2019 to September 2020. They were divided into two groups in accordance with the principle of patient voluntariness: 237 cases in the experimental group and 143 cases in the control group. Patients in both groups were given low-frequency electrical stimulation after surgery. In addition, patients in the experimental group began to take the Dingkun pill orally (one pill per time, two times per day) from the first day of surgery and continued to take it until their menstruation returned to normal. Abdominal pain, the duration of vaginal bleeding, and the amount of bleeding were observed in both groups. Uterine size, endometrial thickness, and urinary human chorionic gonadotropin (HCG) status were also observed at 2 weeks postoperatively to determine preoperative and postoperative HSP70 and HSP90 serum levels. The time of menstrual resumption, menstrual period, and menstrual volume were observed and compared with preoperative menstruation. The occurrence of complications, such as a residual uterine cavity, uterine effusion, menstrual irregularities, and reproductive tract infections, during the follow-up period was also recorded in both groups. Results: Comparison of the endometrial thickness (mm) and uterine size (sum of the three diameters) on uterine adnexal ultrasound at 2-week postoperative review was better in the experimental group than in the control group (p < 0.05). No statistically significant difference was found between the two groups in terms of residual uterine cavity and blood accumulation in the uterine cavity and the results of the urine pregnancy test (p > 0.05). Serum HSP70 and HSP90 levels were significantly higher in the control group than in the experimental group 2 weeks after surgery (p < 0.05). Postoperative HSP70 and HSP90 levels were significantly higher than preoperative levels in both groups (p < 0.05). The degree of postoperative abdominal pain in the experimental group was less severe than that in the control group, and the duration was shorter (p < 0.05). No statistically significant differences were observed when comparing the duration of postoperative vaginal bleeding and the amount of bleeding (p > 0.05). The time of the first menstrual resumption and menstrual volume were more satisfactory in the experimental group than in the control group (p < 0.05). No adverse reactions occurred in either group of patients receiving treatment. Conclusion: Low-frequency electrical stimulation combined with tonifying the kidney and invigorating blood pills could effectively promote uterine rejuvenation after abortion, conducive to improving patients' postoperative abdominal pain, promoting menstrual recovery and maintaining menstrual flow.

Fabrication of One-Dimensional Organic Nanofiber Networks via Electrophoretic Deposition for a Nonvolatile Memory Device.

Despite many advanced growth methodologies for organic nanofibers (ONFs), the lack of efficient and scalable ONF-based film preparation technologies has long been a hindrance in their practical application in organic electronic devices. Here, a typical cathode electrophoretic deposition (C-EPD) technology was developed to controllably produce ONFs and their corresponding thin films. Using the solvent effect and an external electric field force during the C-EPD process, a one-dimensional ONF network was formed, which exhibits compact molecular packing and superior optoelectronic properties in the thin-film state. Prototype sandwich-structure memory devices based on these ONF films exhibited a binary nonvolatile memory performance significantly superior than that of the bulk materials. This study provides an efficient and scalable ONF fabrication technology for high-performance electronic devices in various potential applications.

Effects of naringin on reversing cisplatin resistance and the Wnt/ß-catenin pathway in human ovarian cancer SKOV3/CDDP cells.

OBJECTIVE: Ovarian cancer is one of three malignant tumors of the female reproductive system. Our previous studies showed that the traditional Chinese medicine naringin significantly inhibited the proliferation of platinum-resistant ovarian cancer cells in vitro, and that the mechanism may be related to the NF-κB pathway. METHODS: The MTT assay was used to detect the sensitivity of SKOV3 and SKOV3/CDDP cells to cisplatin, the effect of different naringin concentrations on the proliferation of SKOV3/CDDP cells, and the reversal of cisplatin resistance in naringin-treated SKOV3/CDDP cells. Western blotting was used to detect ß-catenin, c-Myc, and cyclin D1 protein levels in the different cell lines. RESULTS: MTT results showed that different concentrations of naringin inhibited the proliferation of SKOV3 and SKOV3/CDDP cells, and that the inhibition increased with increasing concentrations and prolonged incubation times. Western blotting revealed that compared with controls (SKOV3/CDDP-0), ß-catenin, c-Myc and cyclin D1 proteins levels were significantly decreased in SKOV3/CDDP-C, SKOV3/CDDP-N 20, and SKOV3/CDDP-CN 20 cells, suggesting that naringin inhibited the proliferation of SKOV3/CDDP cells in a concentration and time dependent manner. CONCLUSIONS: Non-cytotoxic naringin reduced the expression of ß-catenin, c-Myc, and cyclin D1 in SKOV3/CDDP cells and partially reversed cisplatin resistance in SKOV3/CDDP CN 20 cells.

Effects of activating GABAB1 receptor on proliferation, migration, invasion and epithelial-mesenchymal transition of ovarian cancer cells.

OBJECTIVE: This study aimed to explore the effects of activating GABAB1 receptor by baclofen on proliferation, migration, invasion and epithelial-mesenchymal transition (EMT) of ovarian cancer cells. RESULTS: One hundred µmol/L, 200 µmol/L and 300 µmol/L were selected as low, medium and high baclofen concentrations respectively. Cells were divided into four groups: Control, 100 µmol/L, 200 µmol/L and 300 µmol/L. Compared with the control group, the viability, colony formation, migration and invasion of SKOV3 cells were inhibited, and the apoptosis of SKOV3 cells were enhanced significantly at 200 µmol/L and 300 µmol/L baclofen. Moreover, they changed significantly with the increase of baclofen concentration. Compared with the control group, the expression of E-cadherin and GABAB1 increased and the N-cadherin expression decreased significantly in 200 µmol/L and 300 µmol/L groups. Higher concentration of baclofen induced higher expression of E-cadherin and lower expression of N-cadherin. CONCLUSION: Baclofen inhibited the proliferation, cloning, migration, invasion and EMT of ovarian cancer cells by activating GABAB1 receptor. These results might contribute a lot to clarify the role and possible mechanism of GABAB1 receptor in ovarian cancer.

Zinc and Cadmium Complexes of Pyridinemethanol Carboxylates: Metal Carboxylate Zwitterions and Metal-Organic Frameworks.

The heterofunctional lactone furo[3,4-b]pyridin-5(7H)-one (L1 ) undergoes a coordination-induced ring-opening reaction with Zn(NO3 )2 ⋅ 6H2 O to yield the zwitterionic [Zn(L1 ')2 (H2 O)2 ] (1, L1 '=2-(hydroxymethyl)nicotinate) with an uncoordinated carboxylate. The same reaction with Cd(NO3 )2 ⋅ 4H2 O provides a two-dimensional (2D) network of [Cd(L1 ')2 ]n (3) with the carboxylates coordinated to cadmium(II) propagating the assembly. The corresponding reactions of Zn(NO3 )2 ⋅ 6H2 O and Cd(NO3 )2 ⋅ 4H2 O with 2-(hydroxymethyl)isonicotinic acid (HL2 ) generated zwitterionic [Zn(L2 )2 (H2 O)2 ] (2) and a 2D network [Cd(L2 )2 ]n ⋅nDMF (4, DMF=N,N'-dimethylformamide), respectively. Complexes 1-4 are weakly emissive, giving ligand-centered emissions at 409 nm (1), 412/436 nm (2), 404 nm (3), and 412/436 nm (4) in CHCl3 solutions upon excitation at 330 nm. This work points to the potential of using 'hidden' functionalities widely found in small organic molecules and natural products for the construction of coordination complexes with new functionality and potential applications.

Systematic Review with Meta-Analysis: The Effects of Probiotics in Nonalcoholic Fatty Liver Disease.

BACKGROUND AND AIMS: Probiotics was considered as a potential therapy for nonalcoholic fatty liver disease (NAFLD) without approval and comprehensive assessment in recent years, which call for a meta-analysis. METHODS: We performed electronic and manual searches including English and Chinese databases published before April 2019, with the use of mesh term and free text of "nonalcoholic fatty liver disease" and "probiotics." Clinical trials evaluating the efficacy of probiotic therapy in NAFLD patients were included according to the eligibility criteria. With the use of random effects models, clinical outcomes were presented as weighted mean difference (WMD) with 95% confidence interval (CI), while heterogeneity and meta-regression were also assessed. RESULTS: 28 clinical trials enrolling 1555 criterion proven NAFLD patients with the use of probiotics from 4 to 28 weeks were included. Overall, probiotic therapy had beneficial effects on body mass index (WMD: -1.46, 95% CI: [-2.44, -0.48]), alanine aminotransferase (WMD: -13.40, 95% CI: [-17.03, -9.77]), aspartate transaminase (WMD: -13.54, 95% CI: [-17.86, -9.22]), gamma-glutamyl transpeptidase (WMD: -9.88, 95% CI: [-17.77, -1.99]), insulin (WMD: -1.32, 95% CI: [-2.43, -0.21]), homeostasis model assessment-insulin resistance (WMD: -0.42, 95% CI: [-0.73, -0.12]), and total cholesterol (WMD: -15.38, 95% CI: [-26.50, -4.25]), but not in fasting blood sugar, lipid profiles, or tumor necrosis factor-alpha. CONCLUSION: The systematic review and meta-analysis support that probiotics are superior to placebo in NAFLD patients and could be utilized as a common complementary therapeutic approach.

Pinocembrin inhibits the proliferation and migration and promotes the apoptosis of ovarian cancer cells through down-regulating the mRNA levels of N-cadherin and GABAB receptor.

There is no previous study on the effect of pinocembrin on ovarian cancer to the best of our knowledge. Moreover, the effects of pinocembrin on the expression of GABAB1 and GABAB2 genes are not studied before. Therefore, this study aimed to investigate effects of pinocembrin on the growth of ovarian cancer cells and the expression of cadherin and GABAB receptor to explore whether pinocembrin was helpful in the treatment of epithelial ovarian cancer. SKOV3 cells were divided into six groups: Control (blank control), DDP (cisplatin as positive control; cells were incubated with 15 µg/ml DDP), 25 µM (cells were incubated with 25 µM pinocembrin), 50 µM (cells were incubated with 50 µM pinocembrin), 100 µM (cells were incubated with 100 µM pinocembrin), and 200 µM (cells were incubated with 200 µM pinocembrin). CCK8 assay, cell scratch assay and Annexin V-FITC/PI staining found that when pinocembrin concentration reached 100 µM and the treatment time reached 48 h, pinocembrin could inhibit the cell proliferation and migration and promote the cell apoptosis, and this effect was enhanced with the increase of pinocembrin concentration. Western blotting found that the protein expression of E-cadherin, N-cadherin, GABAB1 and GABAB2 was not significantly affected by pinocembrin. RT-PCR found that pinocembrin also had no significant influence on the E-cadherin mRNA level, but it could reduce the mRNA levels of N-cadherin, GABAB1 and GABAB2. In conclusion, pinocembrin inhibited the proliferation and migration and promoted the apoptosis of ovarian cancer cells through down-regulating the mRNA levels of N-cadherin and GABAB receptor.

Effects of stocking density on oxidative stress status and mammary gland permeability in early lactating dairy cows.

The current study was conducted to investigate the effect of stocking density (SD) on oxidative stress status and mammary gland permeability in early lactating dairy cows. Thirty-two dairy cows were allocated into 16 blocks, basing on parity, previous milk yield, and body weight, and were then randomly assigned into one of the two treatments as follows: 75% (75SD) and 100% (100SD) SD. The cows were fed with same diet throughout the 8-week experimental period. The milk yield and milk sample were collected on two consecutive days during the 8-week experimental period weekly. Plasma samples were collected on fourth and eighth experimental weeks. Raw, energy-corrected, and 4% fat-corrected milk yield were significantly higher in 75SD-cows than that of 100SD-animals, respectively. The milk somatic cell count was lower in 75SD-cows than that of 100SD-animals. The levels of Na+ , Na+ /K+ , bovine serum albumin and plasmin were lower in 75SD-cows than those of 100SD-cattle, respectively. The 75SD-cows had reduced insulin and insulin/glucagon levels but higher prolactin and growth hormone concentrations, compared with those of 100SD-animals, respectively. In conclusion, compared with low SD-animals, early lactating cows with higher SD had higher oxidative stress status, which further led to a greater mammary gland permeability.

Effective loading of cisplatin into a nanoscale UiO-66 metal-organic framework with preformed defects.

Defects within the nanoscale UiO-66 metal-organic framework (MOF) are created to lock a hybrid phosphonoacetate ligand through Zr-O-P linkages, leaving the carboxyl group free to anchor cisplatin prodrug cis, cis, trans-[Pt(NH3)2Cl2(OH)2]. A drug loading of 256.5 mg g-1 (25.7 wt% based on cisplatin) was achieved with a Zr6 : Pt : P ratio of 1.5 : 1 : 1, which surpasses defect-free UiO-66 and several other MOF carriers. This framework exhibited a burst release of its payload in PBS solution in the first 2 h, releasing 71% of the drug, including a 50% payload release in less than 1 h. This work demonstrates that MOF defects can be intentionally engineered to achieve a high drug loading, and serves as an alternative to drug encapsulation using the pore void and through the association of the functionalized ligand.

Effects of Stocking Density on Milk Fatty Acids Composition and Oxidative Stability of Mid- and Late-Lactating Dairy Cows.

The objective of the current study was to investigate the effects of stocking density (SD) on the milk fatty acid profile and oxidation stability in mid- and late-lactating cows. Twenty-four multiparous mid-lactating Holstein dairy cows (milk yield = 34.5 kg/d (standard deviation 0.68), days in milk = 106 (standard deviation 2.2)) and 24 multiparous late-lactating animals (milk yield = 31.8 kg/d (standard deviation 0.98), DIM = 201 (standard deviation 3.5)) were allocated into 12 blocks based on parity, milk yield, and DIM and were randomly assigned to one of four treatments: mid-lactating cows with high SD (HSD, 100%), mid-lactating cows with low SD (LSD, 75%), late-lactating cows with HSD, and late-lactating cows with LSD. The experiment lasted for eight weeks. Lactation performance (milk yield and composition including fat, protein and lactose) was measured weekly. The milk fatty acids (FAs) profiles and oxidation stability indexes in milk were measured in the fourth day of the eighth week. Plasma variables were measured in the fourth day of fourth and eighth experimental weeks. Yield and composition (protein, fat and lactose) were not affected by SD in neither mid- nor late-lactating dairy cows. Among mid-lactating cows, LSD cows had higher contents of unsaturated FAs (total, C18:1 cis-9 and C18:2 cis-9 cis-12) and lower concentrations of saturated FAs (total, C14:0 and C16:0) in milk fat than those of the HSD animals. Moreover, LSD lowered SOD, GSH-px, and T-AOC activities and reduced the malonaldehyde content in the milk of mid-lactating cows compared with those of HSD cows. Mid-lactating cows under LSD had reduced cortisol and greater MDA contents in plasma than those of HSD cows. Our results suggested that the effect of SD on the milk FA profile and stability varied depending on lactation stages. In mid-lactating cows, although cows with LSD were less stressed, the milk they produced had lower oxidation stability due to the higher unsaturated FAs levels compared with that of HSD cows.

Effect of dietary supplementation of gallic acid on nitrogen balance, nitrogen excretion pattern and urinary nitrogenous constituents in beef cattle.

The objective of the trial was to study the effects of dietary supplementation of gallic acid (GA) on nitrogen (N) balance, N excretion pattern and urinary N constituents in beef cattle. In a 4 × 4 Latin square design, four male 30-month-old Simmental cattle (443 ± 22 kg live weight) received four levels of GA (purity ≥ 98.5%), i.e. 0, 5.3, 10.5, 21.1 g/kg DM, added to a basal ration. Each experimental period lasted 17 d, consisting of 12 d adaptation and 5 d sampling. The results showed that supplementation of GA at 5.3, 10.5 or 21.1 g/kg DM did not affect the N balance but regulated the N excretion pattern by increasing the ratio of faecal N/urinary N and decreasing the ratio of urinary urea N/total urinary N in beef cattle fed at maintenance level.

Supplementing Vitamin E to the Ration of Beef Cattle Increased the Utilization Efficiency of Dietary Nitrogen.

The objectives of the trial were to investigate the effects of supplementing vitamin E (VE) on nutrient digestion, nitrogen (N) retention and plasma parameters of beef cattle in feedlot. Four growing Simmental bulls, fed with a total mixed ration composed of corn silage and concentrate mixture as basal ration, were used as the experimental animals. Four levels of VE product, i.e. 0, 150, 300, 600 mg/head/d (equivalent to 0, 75, 150, 300 IU VE/head/d), were supplemented to the basal ration (VE content 38 IU/kg dry matter) in a 4×4 Latin square design as experimental treatments I, II, III and IV, respectively. Each experimental period lasted 15 days, of which the first 12 days were for pretreatment and the last 3 days for sampling. The results showed that supplementing VE did not affect the nutrient digestibility (p>0.05) whereas decreased the urinary N excretion (p<0.01), increased the N retention (p<0.05) and tended to increase the microbial N supply estimated based on the total urinary purine derivatives (p = 0.057). Supplementing VE increased the plasma concentrations of VE, glucose and triglycerol (TG) (p<0.05) and tended to increase the plasma concentration of total protein (p = 0.096) whereas did not affect the plasma antioxidant indices and other parameters (p>0.05). It was concluded that supplementing VE up to 300 IU/head/d did not affect the nutrient digestibility whereas supplementing VE at 150 or 300 IU/head/d increased the N retention and the plasma concentrations of VE and TG (p<0.05) of beef cattle.

Comparison of the effects of lanthanum, cerium and praseodymium on rumen fermentation, nutrient digestion and plasma biochemical parameters in beef cattle.

The objectives of the trial were to compare the effects of supplementing rare earth elements (REE) lanthanum (La), cerium (Ce) and praseodymium (Pr) on rumen fermentation, nutrient digestion, methane (CH4) production, nitrogen (N) balance and plasma biochemical parameters in beef cattle. Four Simmental male cattle, aged 12 months, with initial average liveweight of 333 ± 9 kg and fitted with rumen cannulas, were fed with a basal ration composed of concentrate mixture and maize silage. Animals received a basal ration without adding REE (Control) or three treatments, i.e. supplementing LaCl3, CeCl3 or PrCl3 at 204 mg/kg DM to the basal ration, respectively, which were allocated in a 4 × 4 Latin square design. Each experimental period lasted 15 d, consisting of 12 d for pre-treatment and three subsequent days for sampling. Results showed that all tested levels of REE tended to increase neutral detergent fibre digestibility (p = 0.064) and tended to decrease rumen CH4 production (p = 0.056). Supplementing LaCl3 and CeCl3 decreased total N excretion and urinary N excretion, increased N retention (p < 0.05), tended to increase total urinary purine derivatives (PD) (p = 0.053) and microbial N flow (p = 0.095), whereas supplementing PrCl3 did not affect N retention, urinary PD and microbial N flow. No differences were found in the effects of nutrient digestibility, CH4 production and plasma biochemical parameters among LaCl3, CeCl3 and PrCl3. Further trials using graded levels of LaCl3, CeCl3 and PrCl3 in a wide range are needed to obtain more pronounced results for comparing effects of La, Ce and Pr on rumen fermentation and nutrient digestion in beef cattle.