Objective To establish a separation method for neoeriocitrin and naringin reference sub- stances from Drynaria fortunei.Methods The extract of D.fortunei was isolated and purified by macro porous resin,silica gel column chromatography,neutro-aluminum oxide adsorbing,Sephadex LH-20 co- lumn chromatography,and recrystallizations.The structures of neoeriocitrin and naringin were identified by MS,IR,UV,~1H-NMR,and ~(13)C-NMR.Results The contents of neoeriocitrin and naringin were 99.5% and 99.3%,respectively.Conclusion The developed method is simple with lower cost,by which neoeriocitrin and naringin can be used as reference substances for the qualitative and quantitative analysis of Chinese herbal medicine.

AIM:To observe the changes in heme oxygenase-1(HO-1) expression in the lung after ischmia-reperfusion of hind limbs in rats.METHODS:Hind limbs ischemia was made by clamping infrarenal aorta with a microvascular clip and lung injury was made by following reperfusion. Lung tissue was obtained from the animals subjected to sham operation, 4 h ischemia without reperfusion and 4 h, 8 h, 16 h, 24 h, 48 h reperfusion following 4 h ischemia. The levels of HO-1 mRNA and protein were measured at different times by Northern blot and Western blot. Immunohistochemistry technique was used to determine the cell types responsible for limb ischemic reperfusion induced HO-1 expression. RESULTS:After ischemia-reperfusion of limbs, HO-1 mRNA increased by 4 h, reached a peak at 16 h, and returned toward baseline at 24-48 h. This time course correlated with increased HO-1 protein. Immunohistochemical studies showed HO-1expressed in a variety of cell types, including the airway epithelium, alveolar macrophages and vascular smooth muscular cells. There were no positive signals in sham group and ischemia group both in mRNA levels and protein levels. CONCLUSION:The expression of HO-1 in the lung is not induced by limb ischemia or sham operation, but induced by limb reperfusion after ischemia in rats.

Objective To investigate the progression of mild cognitive impairment (MCI) to Alzheimer's disease (AD) and the factors influencing the related changes in cognitive ability.Methods Seventy-five subjects with mild cognitive impairment (the MCI group),32 with Alzheimer's disease (the AD group) and 17 others with normal cognition (the NC group) were recruited.The Montreal Cognitive Assessment (MOCA) and the Mini-mental State Examination (MMSE) were used to assess their cognitive ability.At the same time,relevant clinical information such as their general condition and past history of disease were recorded.The subjects were followed up for 20 months on average to evaluate their annual rates of progression (APRs),and logistic regression was used to highlight any influencing factors.Results By the end of the follow-up,9 of the 75 MCI subjects had progressed to AD,with an APR of 5.25％.Thirteen cases had recovered normal cognitive functioning (97.6 per 1,000 person-years).Also,2 cases in the NC group (11.76％) developed MCI (69.1 per 1,000 person-years),but none of them had yet progressed to AD.Both hyperlipidemia and a body mass index (BMI) lower than 24 kg/m2 significantly predicted the deterioration of cognitive functioning.Heart disease was significantly correlated with cognitive improvement,and selfmanagement of cognitive function was also a significant protective factor.Conclusions Patients with MCI are at greater risk of developing AD than normal persons.Prevention and early treatment of hyperlipidemia as well as maintaining a normal BMI may delay the deterioration of cognitive functioning.Self-management of cognitive function can improve cognition.

An investigation of inappropriate feeding behavior and its impact was conducted in patents of 875 infants and toddlers aged 1-24 months from March 2004 to February 2008.The survey revealed that inappropriate feeding behavior included feeding sugar solution or sweet fruits daily in 72.1％ parents (631/875) ; adding yolk as the first supplementary food in 51.3％ (394/768) ; not allowing handgrabbing food or coarse food for infants over 7 months in 50.4％ (270/536) and 44.0％ (236/536) ; no meat eating for infants over 12 months in 36.0％ (54/150).And also revealed that 34.0％ (270/795) and 24.8％ (184/743) parents mixed rice flour or yolk with formula milk,respectively; 17.6％ (48/273) infant over 10 months only had cereal; insufficient or excessive water were used for reconstituting milk powder in 11.8％ (87/735) and 9.3％ (68/735),respectively.The results of investigation indicate that inappropriate feeding behavior among infants and toddlers are quite common and the reasons are various.

Objective To explore the peripheral blood T lymphocytes cell level in different grades of cerebral gliomas pa-tients at the time of before and after operation and its clinical significance .Methods A total of 80 cases of brain tumor patients from February 2010 to February 2012 in this hospital were chosed as study objects ,included 57 cases of glioma ,23 cases of other brain tumors ;57 cases of glioma were divided into 19 cases of low grade group and 38 cases of high grade group accorded to WHO grading standards ,and 50 cases of healthy people in same period were selected as control group .Venous blood in three groups were extracted at 1st day and 1st week after operation ,to detected the level of T lymphocyte subsets in peripheral blood and analyze the relationship between the different grade gliomas and prognosis .Results The peripheral blood CD3 + ,CD4 + ,CD4 + /CD8 + in control group were significantly higher than that of other brain tumor group and the glioma group ,CD8 + was significantly lower than that of the two groups(P< 0 .05) .Compared with other tumor group ,peripheral blood CD3 + ,CD4 + ,CD4 + /CD8 + in glioma group de-creased significantly ,CD8 + increased significantly(P< 0 .05) ;before and after the operation ,the CD3 + and CD4 + /CD8 + levels were significantly higher in low grade group than in the high grade group ,CD8 + was significantly lower in low grade group than in the high grade group(P< 0 .05) ,at the time of 1st week after operation ,CD3 + and CD4 + /CD8 + increased ,CD8 + decreased significant-ly of two groups when intra-group comparison (P < 0 .05) .The median survival time were 31 months in low grade group and 13 months in high grade group ,and the singnificance was found in two groups(P< 0 .05) .The median survival time were 34 months in CD4 + /CD8 + > 1 group and 17 months in CD4 + /CD8 + < 1 group ,the singnificance was found in two groups(P< 0 .05) .Conclusion The immune function of patients with brain glioma is inhibited ,the higher the malignancy of the tumor ,the more obvious of im-mune inhibition ;T lymphocyte subsets level could be used as evaluating index of malignant degree and prognosis of brain glioma .

Objective To explore rules of changes in velocity of ultrasound wave transmission at the tibia and radius by a quantitative bone mineral density ultrasound scanner in examination of early incipient rickets in infants and its significance in clinical diagnosis.Methods One hundred and fifty-seven infants who visited child health-care clinic of the Capital Institute of Pediatrics,Beijing during May 2004 to December 2007 and clinically diagnosed as rickets according to the Criteria of Diagnosis for Rickets in Infants and Young Children formulated in 1986 by the Ministry of Health,as well as 124 normal healthy infants as controls,were enrolled in the study and followed-up for three months.Velocity of ultrasound wave transmission at the tibia and radius in all the infants were measured by a bone mineral density ultrasound scanner (Sunlight Omnisense 7000R made in Israel).Results One hundred and fifty-seven infants were clinically diagnosed as rickets,127 of them undergone with carpal plane roentgenography and 90 of the 127 with positive change in bone x-ray imaging and 37 without it.Velocity of ultrasound wave transmission at the tibia and radius measured by z-score was significantly lower in infants with clinically diagnosed rickets than that in healthy controls (Z-values of-10.411 and-10.399 at the tibia and-5.646 and-5.517 at the radius,respectively,P = 0.000 with Mann-Whitney test).Velocity of ultrasound wave transmission at the tibia and radius measured by z-score was lower in those with positive change in x-my imaging than that in those without it,but not reaching a level of statistically significant difference.Velocity of ultrasound wave transmission and z-score correlated positively with change in bone x-ray imaging,respectively with Spearman coefficients of correlation of 0.581 and 0.677 for tibia,0.316 and 0.467 for radius (P = 0.000).Receiver operating characteristic (ROC) curve was drawn from those with rickets and positive left carpal plane roetgenograph.Area under curve of ROC for z-score of velocity of ultrasound wave transmission at the tibia was 0.812 and 0.799 (95% CI 0.758-0.856 and 0.742-0.855,P =0.000),respectively.Area under ROC curve of z-score of velocity of ultrasound wave transmission at the radius was 0.715 and 0.697 (95% CI 0.650-0.780,0.631-0.764,P =0.000),respectively.Cut-off value of z-score was-205--1.95 at the tibia,according to the largest sensitivity and the least false-positivity,with sensibility of 0.8 to 0.9 and specificity of 0.733 to 0.702.As its sensitivity at the radius was more than 0.7,its specificity was lower than 0.524.Conclusions Quantitative ultrasound scanning can be used to detect insufficient bone mineral density in infants and their early skeletal change by rickets but without change in bone x-ray imaging.Velocity of ultrasound wave transmission at the tibia can be used as an indicator to predict early rickets in infants rather than its severity.

Objective To study the mechanism of antidepressant effect of Baihe Zhimu decoction based on NLRP1 inflammasome.Methods A chronic unpredictable mild stress(CUMS)induced depression mouse model was constructed.C57BL/6J mice were randomly divided into a control group,CUMS group,low-dose Baihe Zhimu decoction group,high-dose Baihe Zhimu decoction group,and MDP group(NLRP1 activator).After continuous administration for 4 weeks,the behavioral characteristic of mice was investigated.The levels of mice hippocampal inflammatory factors IL-1β、IL-6,TNF-α,and neurotransmitters 5-HT,NE,and DA were detected by ELISA.The expression of inflammasome related proteins NLRP1,ASC,and caspase-1,as well as related proteins in BDNF/TrkB and ERK/AKT/mTOR signaling pathway were detected by Western blot.Immunofluorescence was used to detect changes in BDNF expression in the hippocampus of mice.Results Baihe Zhimu decoction inhibited the activation of NLRP1 inflammasomes in the hippocampus of CUMS mice,improved their behavioral characteristics,increased the level of neurotransmitters in the hippocampus,and reduced the inflammatory response.The NLRP1 activator MDP reversed the above effects of Baihe Zhimu decoction.Baihe Zhimu decoction increased BDNF,TrkB expression,and phosphorylation levels of ERK,AKT,and mTOR proteins,which could be reversed by MDP.Conclusion Baihe Zhimu decoction improved depression like behavior in CUMS mice by inhibiting NLRP1 inflammasome activation and hippocampal neuroinflammatory response,and thereby activating BDNF/TrkB and ERK/AKT/mTOR signaling pathway.

BACKGROUND:Ferroptosis is a mode of programmed cell death distinct from apoptosis,necrosis,and other novel cellular deaths,which occurs mainly due to accumulated lipid peroxidation.Ferroptosis has been shown to be involved in the pathological process following subarachnoid hemorrhage.Baicalein,serving as an adept sequestered of iron,evinces its prowess by quelling lipid peroxidative cascades.Nonetheless,the enigma lingers as to whether baicalein possesses the capacity to ameliorate neuronal ferroptosis,elicited in the wake of early brain injury after subarachnoid hemorrhage. OBJECTIVE:To investigate the effect and mechanism of baicalein on neuronal ferroptosis after subarachnoid hemorrhage. METHODS:Primary neuronal cells were extracted from C57BL/6L fetal mice at 16-17 days of gestation.Hemoglobin was used to stimulate primary neuronal cells to simulate an in vitro subarachnoid hemorrhage model.The viability of primary neuronal cells treated with baicalein at concentrations of 5,15,25,50,and 100 μmol/L for 24 hours was detected by CCK-8 assay to determine the optimal concentration of baicalein.Primary neuronal cells were divided into control group,hemoglobin group,and hemoglobin+baicalein group.The levels of reactive oxygen species and malondialdehyde in cells were detected by kits.The mRNA expressions of ferroptosis-related markers PTGS2,SLC7A11,and glutathione peroxidase 4 were detected by RT-PCR.The primary neuronal cells were further divided into control group,SLC7A11 inhibitor Erastin group,hemoglobin group,hemoglobin+baicalein group,and hemoglobin+baicalein+Erastin group.The expression of the ferroptosis related markers SLC7A11 and glutathione peroxidase 4 was detected by western blot assay. RESULTS AND CONCLUSION:(1)Baicalein(25 μmol/L)was selected as the following experimental concentration.(2)Compared with the hemoglobin group,the level of malondialdehyde and the level of reactive oxygen species were significantly decreased(P<0.05)in the hemoglobin+baicalein group.(3)Compared with the hemoglobin group,the mRNA expression of PTGS2 significantly decreased,and the mRNA expression of SLC7A11 and glutathione peroxidase 4 significantly increased(P<0.000 1)in the hemoglobin+baicalein group.(4)SLC7A11 inhibitor Erastin could reverse the baicalin-improved ferroptosis effect to a certain extent(P<0.05).(5)The results showed that baicalein could alleviate the ferroptosis of neuronal cells after subarachnoid hemorrhage through the SLC7A11/GPX4 pathway.

Objective To investigate the regulatory effect of Xuebijing injection on inflammatory reaction during the preservation of isolated empty beating pig hearts with extracorporeal membrane oxygenation.Methods Twelve healthy Guangxi Bama miniature pigs were randomly divided into the Xuebijing group(n=6)and normal saline group(n=6).After the models were established in the Xuebijing group,Xuebijing injection was given at a dose of 5 mL/h through micropump in membrane oxygenator.In the normal saline group,an equivalent amount of 0.9％sodium chloride injection was pumped.Continuous pumping was performed for 8 h in both groups.The time of cardiac resuscitation and perfusion pressure,heart rate,perfusion flow rate after 8 h preservation were recorded in two groups.Pathological and ultrastructural changes of myocardial tissues in the left ventricular wall of hearts with cardiac arrest were observed after 8 h preservation.Serum levels of myocardial injury markers and inflammatory cytokines were detected in two groups at the beginning(T0),2 h(T2),4 h(T4),6 h(T6)and 8 h(T8)after model establishment,respectively.The expression levels of NOD-like receptor protein 3(NLRP3),cysteinyl aspartate specific proteinase-1(Caspase-1),apoptosis-associated speck-like protein containing a CARD(ASC)messenger RNA(mRNA)in myocardial tissues were measured at T0,T2,T4,T6 and T8,respectively.Results There were no significant differences in the time of cardiac resuscitation and perfusion pressure,heart rate,perfusion flow rate after 8 h preservation between two groups(all P＞0.05).Compared with the normal saline group,the levels of lactate dehydrogenase(LDH)at T4,creatine kinase(CK),LDH and α-hydroxybutyrate dehydrogenase(α-HBDH)at T6 and T8,tumor necrosis factor(TNF)-α at T4,T6 and T8,and interleukin(IL)-6,IL-18 and IL-1 β at T0,T2,T4,T6 and T8 were lower,and the mRNA relative expression levels of NLRP3 and Caspase-1 at T2,T4 and T6,and Caspase-1 and ASC at T8 were lower in the Xuebijing group,respectively(all P＜0.05).Hematoxylin-eosin staining and transmission electron microscopy showed that the degree of myocardial injury in the Xuebijing group was slighter than that in the normal saline group.Conclusions Xuebijing injection may effectively mitigate inflammatory response and exert certain myocardial protection effect during the ECMO preservation of isolated empty beating pig hearts.

Lung cancer is the malignant tumor with the highest incidence and mortality rate worldwide.For lung adenocarcinoma,identifying specific gene mutations,fusions,and giving corresponding targeted drugs can greatly improve the survival time of the patients.Among them,anaplastic lymphoma kinase(ALK)fusion occurs in 3％-7％of non-small cell lung cancer(NSCLC).In clinical practice,a variety of detection methods can be used to determine the ALK fusion status,but false negative test results are possible.This paper retrospectively analyzed the diagnosis and treatment of a patient with lung adeno-carcinoma,judged the ALK fusion status by various detection methods.Among them,immunohistochemistry(IHC)(Ventana D5F3),RNA based next-generation sequencing(RNA-based NGS)confirmed positive echinoderm microtubule associated protein like 4(EML4)-ALK fusion,while DNA-based NGS was negative.This paper analyzed the detection methods of ALK fusion,in order to clarify which detection method is the most accurate and simple to choose in different clinical cases and guide the subsequent treatment.