Protective effects of recombinant lactoferrin with different iron saturations on enteritis injury in young mice.

Infant intestinal development is immature and, thus, is vulnerable to bacterial and viral infections, which damage intestinal development and even induce acute enteritis. Numerous studies have investigated that lactoferrin (LF) has protective effects on the intestine and may play a role in preventing intestinal inflammation in infants. Lactoferrin is divided into 2 types, namely apo-LF and holo-LF, depending on the degree of iron saturation, which may affect its bioactivities. However, the role of LF iron saturation in protecting infant intestinal inflammation has not been clearly clarified. Therefore, in this study, young mice models with intestinal damage induced by lipopolysaccharides (LPS) in vivo and primary intestinal epithelial cells in vitro were constructed to enteritis injury in infants for investigation. The apo-LF and holo-LF were subsequently applied to the mouse models to investigate and compare their levels of protection in the intestinal inflammatory injury, as well as to identify which LF was most active. Moreover, the specific mechanism of the LF with optimal iron saturation was further investigated through Western blot assay. Results demonstrated that disease activity index, shortened length of colon tissue, and histopathological score were significantly decreased in the apo-LF group compared with those of the LPS group and the holo-LF group. In the apo-LF group, the concentration of LPS in the intestinal tract and the number of gram-negative bacteria colonies decreased significantly and the expression levels of proinflammatory factors in the colon tissue were downregulated, in comparison with those in the LPS group. The findings of this study thus verify that apo-LF can significantly alleviate enteritis injury caused by LPS, through regulating the PPAR-γ/PFKFB3/NF-κB inflammatory pathway.

Inhibitory effects of lactoferrin on pulmonary inflammatory processes induced by lipopolysaccharide by modulating the TLR4-related pathway.

This study tested the ability of lactoferrin to modulate pulmonary inflammation. To construct in vitro and in vivo inflammatory lung models, cells from the human lung adenocarcinoma cell line (A549) were exposed to lipopolysaccharide (LPS, 1 µg/mL), and mice (CD-1) were intratracheally administered LPS [10 mg/kg of body weight (BW), tracheal lumen injection], respectively. The A549 cells were preincubated with lactoferrin (10 mg/mL), and the mice were intraperitoneally injected with lactoferrin (100 mg/kg of BW), followed by LPS treatment. The concentrations of proinflammatory cytokines (IL-1ß and TNF-α) in culture medium of A549 cells and in bronchoalveolar lavage fluid of the mice were determined using enzyme-linked immunosorbent assays. The toll-like receptor 4-related pathway (TLR4/MyD88/IRAK1/TRAF6/NFκB) was determined at gene and protein expression levels in A549 cells and mouse lung tissue. Results showed that LPS treatment significantly elevated the concentrations of IL-1ß and TNF-α in the A549 cell culture medium and in bronchoalveolar lavage fluid of the mice; it also elevated both the mRNA and protein expressions of TLR4 and the TLR4 downstream factors in A549 cells and mouse lung tissue. Nevertheless, lactoferrin apparently depressed the releases of IL-1ß and TNF-α from A549 cells and lung tissues stimulated by LPS, and significantly suppressed the TLR4 signaling pathway. Lactoferrin also promoted the enhancement of miR-146a expression in A549 cells and mouse lung tissue. Moreover, 100°C heating for 3 min caused total loss of the previously listed bioactivity of lactoferrin. Collectively, we proved that lactoferrin intervened in LPS-induced inflammation in the pulmonary cell model and in the mouse model, through inhibiting the TLR4-related pathway.

[Effects of iASSIST navigation system and personal specific instrument assisted total knee arthroplasty in the treatment of osteoarthritis].

Objective: To compare the application of iASSIST assisted total knee arthroplasty (TKA) and three-dimentional(3D) printing personal specific instrument (PSI) assist TKA in the treatment of osteoarthritis (OA). Methods: Clinical data of 47 patients with OA admitted at Department of Orthopaedic Surgery in Nanjing Medical University Nanjing Hospital between April and September 2016 were retrospectively reviewed, including 20 males and 27 females, aging from 57 to 77 years with mean age of (63.8±8.2) years. They were randomly divided into iASSIST-TKA group (23 patients) and PSI-TKA group (24 patients). The data such as hip knee ankle (HKA) angle, frontal femoral component (FFC) angle, frontal tibial component (FTC) angle, lateral femoral component (LFC) angle, lateral tibial component (LTC) angle, time of operation, post-operative wound drainage, period of hospitalization, visual analog scale (VAS) and Knee Society Score (KSS) at 1 day, 7 days, 14 days, 1 month and 3 months were recorded and compared between the two groups. T test was used to compare measurement data, Fisher exact test and χ(2) test were applied to enumeration data in comparison among groups, and Kruskal-Wallis test was applied to ranked data. Results: The deviation values of HKA, FFC, LFC, FTC and LTC angles were all below 3°(-2° to 2°), and there were no significant difference between iASSIST-TKA group and PSI-TKA group (Z=-0.610 to 0.000, P=0.542 to 1.000). Compared to PSI-TKA group, the time of operation was long((80.7±8.8) minutes vs.(60.2±7.8) minutes), the amount of post-operative wound drainage was increased((210.7±32.1) ml vs.(185.5±30.2)ml) and the period of hospitalization decreased((5.4±2.4) d vs.(6.7±1.6) d) in iASSIST-TKA group, there were significant difference(t=-2.190 to 8.460, P=0.000 to 0.033). There were no significant difference in intra-operative blood drainage((18.4±5.4) ml vs.(17.3±6.2) ml) between the two groups(t=0.650, P=0.521). PSI-TKA group had a superior VAS score(4.8±0.6 vs. 5.5±0.9, 3.6±0.8 vs. 4.3±0.9), KSS clinical score(49.3±5.5 vs. 44.2±6.4, 54.9±4.0 vs. 50.8±4.2) and KSS function score(44.1±2.9 vs. 41.2±3.5, 49.6±3.8 vs. 46.6±3.2) in 1 day and 7 days post-operation(t=-3.420 to 3.150, P=0.001 to 0.007). There were no significant difference in VAS and KSS score in 14 days, 1 month and 3 months post-operation(t=-1.390 to 0.530, P=0.170 to 1.000) between the two groups. Conclusions: The iASSIST-TKA and PSI-TKA can help to make TKA procedure more accurately. iASSIST-TKA may take longer time of operation and have slower recovery, PSI-TKA may need more X-ray input and longer period of hospitalization. The long-term research of both techniques may be valuable for the further clinical usage.

[Chemical studies on the constituents of Phyllanthus urinaria L].

Two new phenolic compounds (crystal VI and crystal IX) have been isolated from Phyllanthus urinaria L. (Euphorbiaceae). Their structures were determined by analysis of their UV, IR, 1H-NMR, 1H-1H COSY, 13C-1H COSY, long range 13C-1H COSY, DEPT, EIMS and HREIMS spectral data. Crystal VI was determined as methyl brevifolincarboxylate. Crystal IX was elucidated as trimethyl ester dehydrochebulic acid. All of the signals of the 13C-NMR of these two new compounds have been assigned mainly according to DEPT, 13C-1H COSY and long range 13C-1H COSY. Accompanying these two new compounds, 8 known compounds have been isolated. By using chemical reactions, UV, IR, 1H-NMR, MS, their structures were elucidated as n-octadecane(I), beta-sitosterol(III), ellagic acid(IV), daucosterol(V), kaempferol(VII), quercetin(VIII), gallic acid(X) and rutin(XI). Crystal II is an aldehyde, its structure elucidation is in progress.

[Biotransformation of (+)- and (-)-clausenamide in rats].

AIM: To study the metabolic pathway of chiral clausenamide in the rat and understand its stereoselectivity. METHODS: The urine, feces and blood of rat were gathered after the drug was administered, the known metabolites were analyzed by HPLC-DAD and one unknown metabolite was elucidated by using LC-MS analysis. Metabolic stereoselectivity was determined by comparing the metabolic results of (+)- and (-)-clausenamide. RESULTS: Six known metabolites were determined and one unknown metabolite was elucidated as N-demethylclausenamide. The metabolic stereoselectivity was shown distinctly. CONCLUSION: Chiral clausenamide was mainly metabolized by hydroxylation in liver and the biotransformation exhibited pronounced substrate stereoselectivity.