Objective The aim of this study was to establish a reasonable protocol for a rat model of ferric chloride-induced carotid arterial thrombosis by ultrasonic continuous evaluation.Methods Twenty SD rats were randomly divided into four groups and then treated with 20％,30％,40％ or 50％ concentrations of FeCl3,respectively.The vascular conditions were evaluated by 14L ultrasonic probe,at 10 mins,15 mins,and 20 mins.We then selected the best group through the examination of the rate of spontaneous reperfusion of blood vessels and the rate of reperfusion after intravenous injection of urokinase.At the end of the experiment,vessels were fixed in 10％ formalin solution and stained with HE.Results After external application of FeCl3 on rat common carotid artery for 20 mins,the artery occlusion rate was 100％,20％,0％ and 0％ in animals receiving 50％,40％,30％ and 20％ FeCl3,respectively.After external application of FeCl3 on rat common carotid artery for 120 mins,the spontaneous revascularization rate was 0％ in 50％ concentration group whereas were 100％ in rest other groups (P＜ 0.001).In 50％ concentration group,the partial recanalization rate was 40％ after intravenous injection of urokinase.HE staining revealed that the thrombus was dense and the lumen was partially recanalized after the urokinase intervention in 50％ concentration group.Conclusion By use of uultrasonic continuous evaluation of ferric chloride-induced thrombosis of rat common carotid artery,we have demonstrated that external application of 50％ ferric chloride solution for 20 mins is effective for the formation of thrombus model,which may be suitable for the studv of thrombolysis.

OBJECTIVETo screen a new strain which can transform panaxadiol saponins into the rare ginsenoside compound K.

METHODThe total saponins in stems and leaves of Panax notoginseng was used as a substrate in the liquid state fermentation process, and the results were detected by TLC and HPLC-ELSD to screen a strain from twelve plant pathogenic fungi which can produce ginsenoside compound K.

RESULTFusarium moniliforme was found to transform the total saponins to ginsenoside compound K efficiently in the all twelve fungal strains. In the fermentation process, ginsenoside Rb1 was transformed almost completely, and the content of ginsenoside Rd was decreasing evidently.

CONCLUSIONF. moniliforme is selected as a new high-yield strain. It is expected to be used to produce the high activity infrequent ginsenoside compound K and to improve the content of active principles in medicinal plants.

Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Fungi ; pathogenicity ; Ginsenosides ; chemistry ; Panax notoginseng ; chemistry ; microbiology

Objective To investigate the effects of ω-3 polyunsaturated fatty acids(ω-3PUFAs)and ω-6 polyunsaturated fatty acids(ω-6PUFAs)on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway,and the expressions of tumor necrosis factor-α(TNF-α),interleukin(IL)-1β and IL-6 in neonatal rats with brain injury induced by lipopolysaccharide (LPS). Methods Ninety-six neonatal rats were divided into control group,ω-3PUFAs group,ω-6PUFAs group,and LPS group by using random number table method. Intraperitoneal injection of LPS was performed in LPS group,ω-6PUFAs group and ω-3PUFAs group to establish models of rat brain injury. The rats in control group received 9 g/L saline. Twelve newborn rats were killed at 1 d or 5 d after intraperito-neal injection in each group for hippocampus selection. Real -time PCR and Western blot were used to detect the mRNA and protein expression levels of TLR4,NF-κB,TNF-α,IL-1β and IL-6. Results One day after mode-ling,TLR4,NF-κB,TNF-α,IL-1β and IL-6 mRNA expressions in ω-3PUFAs group (10. 63 ± 0. 07,5. 86 ± 1. 05,7. 65 ± 2. 29,5. 23 ± 1. 31,3. 36 ± 0. 72)were lower than those in ω-6PUFAs group (18. 83 ± 2. 10,8. 79 ± 2. 08,11. 95 ± 3. 23,10. 97 ± 2. 24,6. 37 ± 1. 17)and LPS group (15. 76 ± 1. 59,7. 13 ± 1. 10,9. 71 ± 2. 14,7. 83 ± 0. 85,4. 78 ± 0. 51),and the differences were all statistically significant(all P＜0. 05);which in ω-6PUFAs group were higher than those in LPS group,and the differences were all significant (all P＜0. 05). TLR4,NF-κB,TNF-α, IL-1β and IL-6 protein levels in ω-3PUFAs group (1. 57 ± 0. 11,1. 58 ± 0. 09,1. 55 ± 0. 09,1. 63 ± 0. 31,1. 36 ± 0. 12)were lower than those in ω-6PUFAs group (1. 96 ± 0. 17,2. 21 ± 0. 12,1. 95 ± 0. 23,1. 97 ± 0. 24,1. 77 ± 0. 17)and LPS group (1. 73 ± 0. 15,1. 87 ± 0. 10,1. 79 ± 0. 14,1. 83 ± 0. 15,1. 58 ± 0. 11)in 1 d,and the diffe-rences were all significant (all P＜0. 05),and those in ω-6PUFAs group were higher than those in LPS group (all P＜0. 05). Similarly,TLR,NF-κB,TNF-α,IL-1β and IL-6 mRNA and protein expression levels in ω-3PUFAs group (3. 78 ± 0. 88,3. 86 ± 0. 62,6. 26 ± 1. 94,3. 65 ± 1. 44,2. 11 ± 0. 87;1. 15 ± 0. 08,1. 32 ± 0. 10,1. 46 ± 0. 04, 1. 38 ± 0. 14,1. 21 ± 0. 09)were lower than those in ω-6PUFAs group (7. 76 ± 1. 65,5. 51 ± 0. 88,7. 96 ± 2. 13,5. 35 ± 1. 75,4. 88 ± 1. 35;1. 42 ± 0. 15,1. 51 ± 0. 36,1. 65 ± 0. 13,1. 72 ± 0. 23,1. 48 ± 0. 10)and LPS group (6. 21 ± 1. 87, 4. 98 ± 0. 73,7. 11 ± 2. 10,4. 84 ± 1. 75,4. 25 ± 0. 64;1. 35 ± 0. 13,1. 44 ± 0. 22,1. 59 ± 0. 10,1. 61 ± 0. 18,1. 35 ± 0. 07) in 5 d (all P＜0. 05),and which in ω-6PUFAs group were higher than those in LPS group,and the differences were sig-nificant (all P＜0. 05). Conclusion ω-6PUFAs can up-regulate the activity of TLR4,NF-κB,and reduce the re-lease of TNF-α,IL-1β and IL-6;and ω-3PUFAs can down-regulate the activity of TLR4,NF-κB,and reduce the release of TNF-α,IL-1β and IL-6,so it has a neural protective effect in brain injury induced by LPS.

BACKGROUND/OBJECTIVES: Previous research has shown maternal betaine supplementation alleviates fetal-derived hepatic steatosis. Therefore, this study examined the anti-inflammatory effect of maternal betaine intake in offspring mice and its mechanism.MATERIALS/METHODS: Female C57BL/6J mice and their offspring were randomly divided into 3 groups according to the treatment received during gestation and lactation: control diet (CD), fatty liver disease (FLD), and fatty liver disease + 1% betaine (FLD-BET). The FLD group was given a high-fat diet and streptozotocin (HFD + STZ), and the FLD-BET group was treated with HFD + STZ + 1% betaine. After weaning, the offspring mice were given a normal diet for 5 weeks and then dissected to measure the relevant indexes. RESULTS: Compared to the CD group, the offspring mice in the FLD group revealed obvious hepatic steatosis and increased serum levels of alanine aminotransferase, interleukin (IL)-6, and tumor necrosis factor (TNF)-α; maternal betaine supplementation reversed these changes. The hepatic mRNA expression levels of IL-6, IL-18, and Caspase-1 were significantly higher in the FLD group than in the CD group. Maternal betaine supplementation reduced the expression of IL-1β, IL-6, IL-18, and apoptosis-associated speck-like protein containing C-terminal caspase recruitment domain (ASC). Maternal betaine supplementation also reversed the increasing protein expressions of nitric oxide dioxygenase-like receptor family pyrin domain containing 3 (NLRP3), ASC, Caspase-1, IL-1β, and IL-18 in offspring mice exposed to HFD + STZ. Maternal betaine supplementation decreased the homocysteine (Hcy) and s-adenosine homocysteine (SAH) levels significantly in the livers. Furthermore, the hepatic Hcy concentrations showed significant inverse relationships with the mRNA expression of TNF-α, NLRP3, ASC, and IL-18. The hepatic SAH concentration was inversely associated with the IL-1β mRNA expression. CONCLUSIONS The lipotropic and anti-inflammatory effect of maternal betaine supplementation may be associated with the inhibition of NLRP3 inflammasome in the livers of the offspring mice.

Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.

The correct differentiation of oligodendrocyte precursor cells (OPCs) is essential for the myelination and remyelination processes in the central nervous system. Determining the regulatory mechanism is fundamental to the treatment of demyelinating diseases. By analyzing the RNA sequencing data of different neural cells, we found that cyclin-dependent kinase 18 (CDK18) is exclusively expressed in oligodendrocytes. In vivo studies showed that the expression level of CDK18 gradually increased along with myelin formation during development and in the remyelination phase in a lysophosphatidylcholine-induced demyelination model, and was distinctively highly expressed in oligodendrocytes. In vitro overexpression and interference experiments revealed that CDK18 directly promotes the differentiation of OPCs, without affecting their proliferation or apoptosis. Mechanistically, CDK18 activated the RAS/mitogen-activated protein kinase kinase 1/extracellular signal-regulated kinase pathway, thus promoting OPC differentiation. The results of the present study suggest that CDK18 is a promising cell-type specific target to treat demyelinating disease.

Objective: To investigate the feasibility of long noncoding RNA (lncRNA)_AK096792 as a clinical predictor of bronchopulmonary dysplasia (BPD) in preterm infants. Methods: All the cord blood(2-5 mL) of very low birth weight (VLBW) preterm infants born in Huai′an First Hospital Affiliated to Nanjing Medical University were collected from December 1, 2015 to December 1, 2017.Moreover, the peripheral blood(2 mL) of those VLBW infants diagnosed with BPD was also collected.A total of 36 infants with BPD were collected.Another 36 cases of premature children with VLBW were chosen as control group according to random number table.The relative content of lncRNA_AK096792 in cord blood and peripheral blood was detected by using real-time quantitative PCR (qPCR). Additionally, the correlation of lncRNA_AK096792 levels between cord and peripheral blood of BPD infants was analyzed.The sensitivity and specificity of lncRNA_AK096792 for BPD were analyzed by using receiver operating curve test. Results: (1)LncRNA_AK096792 was a common, evolutionarily conserved, non-coding RNA present in both mouse and human.(2) The expression level of lncRNA_AK096792 in peripheral blood was significantly higher than that in cord blood in BPD group[(463.3±352.0)% vs.(50.0±37.5)%], and the difference was significant(P<0.001), and they were highly correlated (r=0.825, P<0.001). (3) The level of lncRNA_AK096792 in cord blood in BPD group was signi-ficantly higher than that in non-BPD group [(484.3±280.5)% vs.(101.2±28.6)%], and the difference was significant(P<0.001). (4)When lncRNA_AK096792 served as a clinical predictor for BPD, the specificity was 83.3%, the sensitivity was 75.6%, and an area under the receiver operating characteristic curve was 0.88(P<0.001). Conclusions LncRNA_AK096792 is highly correlated with the development of BPD.The level of lncRNA_AK096792 in umbilical cord blood of premature infants can be used as an early predictive marker for BPD, it calls for further study.