MicroRNA-22-3p Regulates the Expression of Kruppel-like Factor 6 to Affect the Cardiomyocyte-like Differentiation of Bone Marrow Mesenchymal Stem Cell / 中国医学科学院学报

Objective To explore the effect of microRNA-22-3p (miR-22-3p) regulating the expression of Kruppel-like factor 6 (KLF6) on the cardiomyocyte-like differentiation of bone marrow mesenchymal stem cell (BMSC). Methods Rat BMSC was isolated and cultured,and the third-generation BMSC was divided into a control group,a 5-azacytidine(5-AZA)group,a mimics-NC group,a miR-22-3p mimics group,a miR-22-3p mimics+pcDNA group,and a miR-22-3p mimics+pcDNA-KLF6 group.Real-time fluorescent quantitative PCR (qRT-PCR) was carried out to determine the expression of miR-22-3p and KLF6 in cells.Immunofluorescence staining was employed to detect the expression of Desmin,cardiac troponin T (cTnT),and connexin 43 (Cx43).Western blotting was employed to determine the protein levels of cTnT,Cx43,Desmin,and KLF6,and flow cytometry to detect the apoptosis of BMSC.The targeting relationship between miR-22-3p and KLF6 was analyzed by dual luciferase reporter gene assay. Results Compared with the control group,5-AZA up-regulated the expression of miR-22-3p (q=7.971,P<0.001),Desmin (q=7.876,P<0.001),cTnT (q=10.272,P<0.001),and Cx43 (q=6.256,P<0.001),increased the apoptosis rate of BMSC (q=12.708,P<0.001),and down-regulated the mRNA (q=20.850,P<0.001) and protein (q=11.080,P<0.001) levels of KLF6.Compared with the 5-AZA group and the mimics-NC group,miR-22-3p mimics up-regulated the expression of miR-22-3p (q=3.591,P<0.001;q=11.650,P<0.001),Desmin (q=5.975,P<0.001;q=13.579,P<0.001),cTnT (q=7.133,P<0.001;q=17.548,P<0.001),and Cx43 (q=4.571,P=0.037;q=11.068,P<0.001),and down-regulated the mRNA (q=7.384,P<0.001;q=28.234,P<0.001) and protein (q=4.594,P=0.036;q=15.945,P<0.001) levels of KLF6.The apoptosis rate of miR-22-3p mimics group was lower than that of 5-AZA group (q=8.216,P<0.001).Compared with the miR-22-3p mimics+pcDNA group,miR-22-3p mimics+pcDNA-KLF6 up-regulated the mRNA(q=23.891,P<0.001) and protein(q=13.378,P<0.001)levels of KLF6,down-regulated the expression of Desmin (q=9.505,P<0.001),cTnT (q=10.985,P<0.001),and Cx43 (q=8.301,P<0.001),and increased the apoptosis rate (q=4.713,P=0.029).The dual luciferase reporter gene experiment demonstrated that KLF6 was a potential target gene of miR-22-3p. Conclusion MiR-22-3p promotes cardiomyocyte-like differentiation of BMSC by inhibiting the expression of KLF6.

An integrated strategy for comprehensive characterization of metabolites and metabolic profiles of bufadienolides from Venenum Bufonis in rats / 药物分析学报

Comprehensive characterization of metabolites and metabolic profiles in plasma has considerable sig-nificance in determining the efficacy and safety of traditional Chinese medicine(TCM)in vivo.However,this process is usually hindered by the insufficient characteristic fragments of metabolites,ubiquitous matrix interference,and complicated screening and identification procedures for metabolites.In this study,an effective strategy was established to systematically characterize the metabolites,deduce the metabolic pathways,and describe the metabolic profiles of bufadienolides isolated from Venenum Bufonis in vivo.The strategy was divided into five steps.First,the blank and test plasma samples were injected into an ultra-high performance liquid chromatography/linear trap quadrupole-orbitrap-mass spectrometry(MS)system in the full scan mode continuously five times to screen for valid matrix compounds and metabolites.Second,an extension-mass defect filter model was established to obtain the targeted precursor ions of the list of bufadienolide metabolites,which reduced approximately 39％of the interfering ions.Third,an acquisition model was developed and used to trigger more tandem MS(MS/MS)fragments of precursor ions based on the targeted ion list.The acquisition mode enhanced the acquisition capability by approximately four times than that of the regular data-dependent acquisition mode.Fourth,the acquired data were imported into Compound Discoverer software for identification of metabolites with metabolic network prediction.The main in vivo metabolic pathways of bufadienolides were elucidated.A total of 147 metabolites were characterized,and the main biotransformation reactions of bufadienolides were hydroxylation,dihydroxylation,and isomerization.Finally,the main prototype bufadienolides in plasma at different time points were determined using LC-MS/MS,and the metabolic profiles were clearly identified.This strategy could be widely used to elucidate the metabolic profiles of TCM preparations or Chinese patent medicines in vivo and provide critical data for rational drug use.

Deep chemical identification of phytoecdysteroids in Achyranthes bidentata Blume by UHPLC coupled with linear ion trap-Orbitrap mass spectrometry and targeted isolation / 中国天然药物

Achyranthes bidentata Blume is widely used as a traditional Chinese medicine with the effects of nourishing the liver and kidneys and strengthening muscles and bones. In this work, a rapid and simple strategy was developed for characterizing phytoecdysteroids by ultra-high-performance liquid chromatography coupled with liner ion trap-Orbitrap mass spectrometry using electrospray ionization in the negative mode. As a result, 47 phytoecdysteroids were unambiguously or tentatively characterized. Among them, seven known compounds were identified according to the reference standards along with molecular formula, retention time and fragmentation patterns, while others were mostly potential new compounds. Through targeted isolation, the structures of three new compounds were determined by NMR spectra, which were consistent with LC-MS characterization. The present study provides an efficient method to deeply characterize phytoecdysteroids.

Identification of chemical constituents of Xiaochengqi Decoction by UPLC-Q-TOF/Fast DDA combined with UNIFI software / 中国中药杂志

Based on the combination of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF) and Waters UNIFI software, the chemical constituents of the classic prescription Xiaochengqi Decoction were qualitatively analyzed and identified. The UPLC conditions are as follows: Acquity HSS T3 reverse phase column(2.1 mm ×100 mm, 1.8 μm), column temperature of 30 ℃, mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B), and flow rate of 0.3 mL·min~(-1). High-resolution MS data of Xiaochengqi Decoction were collected in ESI~(+/-) modes by Fast DDA. The structures of the chemical constituents were tentatively characterized or identified by UNIFI software according to the retention time of reference standards and characteristic fragment ions in MS profile, and literature data. A total of 233 components in Xiaochengqi Decoction were identified, with 93 from wine-processed Rhei Radix et Rhizoma, 104 from bran-processed Aurantii Fructus Immaturus, and 36 from ginger-processed Magnoliae Officinalis Cortex. These 233 components included anthraquinones, flavonoids, lignans, alkaloids, coumarins, and phenylethanoid glycosides. The result provided experimental evidence for the further study on establishment of quality standard and product development of the formula.

Analysis of chemical constituents in classical prescription Danggui Buxue Decoction based on RP-Q-TOF-MS and HILIC-Q-TOF-MS / 中国中药杂志

The chemical constituents of classical prescription Danggui Buxue Decoction were analyzed by reversed-phase(RP) chromatography and hydrophilic interaction chromatography(HILIC) coupled with quadrupole time-of-flight mass spectrometry. RP separation of Danggui Buxue Decoction was performed on ACQUITY UPLC HSS T3(2.1 mm×100 mm, 1.8 μm), while HILIC separation was on Waters BEH Amide(2.1 mm×100 mm, 1.7 μm). Mass spectrometry(MS) data were acquired in both negative and positive ion modes. Chemical constituents of Astragali Radix and Angelicae Sinensis Radix were searched from Reaxys and thus the in-house library was established. MS data were further analyzed by MassLynx 4.1 combined with in-house library, HMDB, Reaxys, and comparison with reference substances. In conclusion, a total of 154 compounds were identified and characterized: 16 saponins, 44 flavonoids, 10 phthalides, 7 phenylpropanoids, 15 bases and the corresponding nucleosides, 30 oligosaccharides, and 32 other compounds. Among them, 65 compounds were detected by HILIC-MS/MS. This study provides experimental evidences for the material basis research, quality control, and preparation development of Danggui Buxue Decoction and a reference method for comprehensive characterization of Chinese medicine decoctions typified by classical prescriptions.

Effect and Mechanism of Banxia Baizhu Tianmatang on Atherosclerosis in ApoE-/- Mice / 中国实验方剂学杂志

Objective:To study the effects of Banxia Baizhu Tianmatang （BBTT） on atherosclerosis in apolipoprotein-E knockout （ApoE^-/-） mice induced by high fat diet. Method:The atherosclerosis model of ApoE^-/- mice was established with high-fat diet， and BBTT was used for intervention. The pathological changes of aorta after atherosclerosis were observed by hematoxylin-eosin （HE）， oil red O and Masson staining. The changes of serum total cholesterol （TC）， triglyceride （TG）， high density lipoprotein cholesterol （HDL-C） and low density lipoprotein cholesterol （LDL-C） were detected by automatic biochemical analyzer. The expression levels of tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-6 （IL-6） and oxidized low density lipoprotein （ox-LDL） were detected by enzyme linked immunosorbent assay （ELISA）. Total tissue proteins were extracted， quantified by protein quantification （BCA） method， and the expression of matrix metalloproteinase-9 （MMP-9） protein was detected by Western blot. Thiobarbituric acid （TBA） method was used to detect the change of malondialdehyde （MDA） content. The change of superoxide dismutase （SOD） activity was detected by 2-（2-methoxy-4-nitrophenyl）-3-（4-nitrophenyl）-5-（2，4-disulfobenzene）-2H tetrazole monosodium salt （WST-8） method. Result:Compared with the control group， there was a large amount of lipid accumulation in the blood vessels of the model group， the serum levels of TG， TC， HDL-C and LDL-C significantly increased （<italic>P</italic><0.05）， the expression of MMP-9 protein in the blood vessels significantly increased （<italic>P</italic><0.01）， the expression levels of IL-6 and TNF-<italic>α</italic> in the serum increased （<italic>P</italic><0.05， <italic>P</italic><0.01）， the SOD activity was significantly reduced （<italic>P</italic><0.01）， and the levels of MDA and ox-LDL expression increased （<italic>P</italic><0.01）. Compared with the model group， the treatment with BBTT could inhibit the accumulation of lipids in blood vessels， the TG levels were reduced in the high and medium dose groups of BBTT （<italic>P</italic><0.05）， high， medium and low dose groups significantly reduced the levels of LDL-C in serum （<italic>P</italic><0.01）， the expression of MMP-9 protein in blood vessels （<italic>P</italic><0.05） and IL-6 in serum （<italic>P</italic><0.01）， the high-dose group down-regulated the expression of TNF-<italic>α</italic> in serum （<italic>P</italic><0.01） and ox-LDL （<italic>P</italic><0.01）， both the high and medium-dose groups increased the level of MDA （<italic>P</italic><0.05， <italic>P</italic><0.01） and the activity of SOD （<italic>P</italic><0.05）. Conclusion:BBTT has a certain intervention effect on the formation of atherosclerosis aortic plaque in ApoE^-/- mice， and its mechanism may be associated with reducing the TG and LDL-C levels， lowering blood lipid， down-regulating MMP-9 protein， protecting blood vessels from inflammatory damage， reducing ox-LDL and MDA levels， and improving SOD activity to play an antioxidant role.

Chemical profiling of Huashi Baidu prescription, an effective anti-COVID-19 TCM formula, by UPLC-Q-TOF/MS / 中国天然药物

Huashi Baidu prescription (HSBDF), recommended in the Guideline for the Diagnosis and Treatment of Novel Coronavirus (2019-nCoV) Pneumonia (On Trials, the Seventh Edition), was clinically used to treat severe corona virus disease 2019 (COVID-19) with cough, blood-stained sputum, inhibited defecation, red tongue etc. symptoms. This study was aimed to elucidate and profile the knowledge on its chemical constituents and the potential anti-inflammatory effect in vitro. In the study, the chemical constituents in extract of HSBDF were characterized by UPLC-Q-TOF/MS in both negative and positive modes, and the pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assays (ELISA) to determine the effects of HSBDF in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The results showed that a total of 217 chemical constituents were tentativedly characterized in HSBDF. Moreover, HSBDF could alleviate the expression levels of IL-6 and TNF-α in the cell models, indicating that the antiviral effects of HSBDF might be associated with regulation of the inflammatory cytokines production in RAW264.7 cells. We hope that the results could be served as the basic data for further study of HSBDF on anti-COVID-19 effect.

Profiling and identification of aqueous extract of Cordyceps sinensis by ultra-high performance liquid chromatography tandem quadrupole-orbitrap mass spectrometry / 中国天然药物

Characterization of aqueous extract in traditional Chinese medicine (TCM) is challenging due to the poor retention of the analytes on conventional C columns. This study presents a systematic characterization method based on a rapid chromatographic separation (8 min) on a polar-modified C (Waters Cortecs T3) column of aqueous extract of Cordyceps sinensis. UHPLC-HRMS method was used to profile components in both untargeted and targeted manners by full MS/PIL/dd-MS acquisition approach. The components were identified or tentatively identified by reference standards comparison, fragmentation rules elucidation and available databases search. A total of 91 components, including 10 nucleobases, 20 nucleosides, 39 dipeptides, 18 amino acids and derivatives and 4 other components, were characterized from the aqueous extract of C. sinensis. And this was the first time to systematically report the presence of nucleosides and dipeptides in C. sinensis, especially for modified nucleosides. The chemical basis inquiry of this work would be beneficial to mechanism exploration and quality control of C. sinensis and related products. Meanwhile, this work also provided an effective solution for characterization of aqueous extract in TCM.

Key factors affecting the implementation of clinical pathways: a systematic review / 中华医院管理杂志

Objective@#To systematically review the barriers and facilitators affecting the implementation of clinical pathways for the clinical pathways.@*Methods@#PubMed, Embase, CNKI, CBM, Wanfang, Cvip databases were searched to collect articles about clinical pathways implementation barriers and facilitators from inception to January 4th, 2019. The tool of confidence in the evidence from reviews of qualitative research(CERQual)was used to grade the confidence of each study.@*Results@#A total of 43 articles from 12 countries were included.There were 8 main categories and 31 subcategories of the barriers about clinical pathways, including content of the clinical pathways, negative outcomes of clinical pathways, physicians knowledge, physicians attitude, resource availability, implementation of activities, patients factors and social factors. The first three barriers of high confidence were lacking of time, capital, equipment, staff and other resources(15 articles, 34.9%), increasing workload(14 articles, 32.6%), unrecognizing pathways(12 articles, 27.9%). There were 6 main categories and 28 subcategories of the facilitators about clinical pathways, including pathways content related, physician related, resource factor and implementation activity. The first three facilitators of high confidence were communication, education and training(25 articles, 58.1%), supporting from managers and colleagues(21 articles, 48.8%)and establishing a clinical pathway facilitation committee(17 articles, 39.5%).@*Conclusions@#The successful implementation of clinical pathways connects with its development process, aftereffect evaluation and feedback. It will be implemented effectively only by the completely and environmentally acceptable pathways design, adequate resources, effective organizational activities, continuous audit, evaluation and feedback and physicians active cooperation.

Key factors affecting the implementation of clinical pathways: a systematic review / 中华医院管理杂志

Objective To systematically review the barriers and facilitators affecting the implementation of clinical pathways for the clinical pathways.Methods PubMed, Embase, CNKI, CBM, Wanfang, Cvip databases were searched to collect articles about clinical pathways implementation barriers and facilitators from inception to January 4th, 2019. The tool of confidence in the evidence from reviews of qualitative research ( CERQual) was used to grade the confidence of each study. Results A total of 43 articles from 12 countries were included.There were 8 main categories and 31 subcategories of the barriers about clinical pathways, including content of the clinical pathways, negative outcomes of clinical pathways, physicians knowledge, physicians attitude, resource availability, implementation of activities, patients factors and social factors. The first three barriers of high confidence were lacking of time, capital, equipment, staff and other resources (15 articles, 34.9% ), increasing workload (14 articles, 32.6% ), unrecognizing pathways(12 articles, 27.9% ). There were 6 main categories and 28 subcategories of the facilitators about clinical pathways, including pathways content related, physician related, resource factor and implementation activity. The first three facilitators of high confidence were communication, education and training(25 articles, 58.1% ), supporting from managers and colleagues ( 21 articles, 48.8% ) and establishing a clinical pathway facilitation committee(17 articles, 39.5% ).Conclusions The successful implementation of clinical pathways connects with its development process, aftereffect evaluation and feedback. It will be implemented effectively only by the completely and environmentally acceptable pathways design, adequate resources, effective organizational activities, continuous audit, evaluation and feedback and physicians active cooperation.

Effects of Fengshi Bitong Prescription on Cytokines in Serum and Synovial Fas/FasL mRNA Expression in Rats with Collagen-Induced Arthritis / 中国中医药信息杂志

Objective To observe the effects of Fengshi Bitong Prescription (FSBT) on cytokines in serum and Fas/FasL mRNA expression system in synovium; To discuss relevant mechanism of action. Methods Rats were given foot intradermal injection of bovine type Ⅱ collagen emulsion to establish collagen-induced arthritis (CIA) model. After successful modeling, they were randomly divided into 6 groups: normal control group, CIA model group, tripterygium glycosides group, FSBT high-, medium-, and low-dose groups. Each medication group was given relevant medicine for gavage for 14 d. The weight and arthritis scores of CIA rats were observed. Serum levels of IL-1β, IL-15, and TGF-β1 were detected by ELISA. Expressions of Fas and FasL mRNA in synovium of CIA rats were detected by real-time PCR. Results Compared with model group, the foot arthritis score and serum levels of IL-15 in FSBT high- and medium-dose groups significantly decreased (P<0.01), and the serum level of TGF-β1 significantly increased (P<0.05). Expression of Fas significantly decreased and FasL significantly increased in FSBT high-dose group (P<0.05). Conclusion FSBT may have certain immune regulation effects on rheumatoid arthritis. Its mechanism may be in regulating Fas/FasL apoptosis system, thereby inducing apoptosis of synovial cells and inhibiting synovial hyperplasia.

Observation on the efficacy of Conbercept for chronic central serous chorioretinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the efficacy of intravitreal conbercept injection for chronic central serous chorioretinopathy (CSC).METHODS: Nine eyes of 9 patients diagnosed as chronic CSC between October 2015 to May 2016 were treated with an intravitreal injection of conbercept (0.5mg/0.05mL) (six patients were given the same does of intravitreal injection again at 1mo after the first injection).Follow-up observation was at 1, 2, and 6mo after injection.Observed indicators included best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), fundus fluorescein angiography (FFA), choroidal indocyanine green angiography (ICGA), macular fovea thickness (CMT), subfoveal choroidal thickness (SFCT).RESULTS:Seven of the 9 patients responded significantly to the drug, while 2 patients had no response.The CMT was 373.12±72.43μm at baseline, which decreased significantly to 332.05±67.13μm, 282.24±62.30μm and 225.56±71.08μm at 1, 2 and 6mo after the intravitreal injection.The mean thickness of SFCT was 422.11±64.82μm before treatment.The choroidal thickness of non-responsive patients before treatment was below average, respectively 353μm and 365μm.The SFCT of 1, 2, and 6mo after treatment was 391.45±75.24μm, 365.53±63.07μm, 355.40±66.65μm.Before treatment and 1mo after, there was no significant difference (P=0.074), but there was statistically significant (P0.05).CONCLUSION: Intravitreal conbercept injection in chronic CSC may have some effect in accelerating subertinal fluid resolution and decreasing the CMT.The SFCT within 6mo after treatment was significantly lower than pretreatment.The SFCT may be an indicator of whether patients respond.

Effect and mechanism of total flavone of epimedium on primary callus formation in ovariectomized rats with fractures / 中国骨伤

<p><b>OBJECTIVE</b>To explore the effects and related mechanisms of total flavone of epimedium treatment(TFE)on primary callus for mation in ovariectomized rats.</p><p><b>METHODS</b>Forty male SD rats weighted from 209 to 246 g and aged 6 to 8 weeks were selected. Six weeks after ovariectomy a femur fracture model with middiaphyseal segment fracture was established, estimated and randomly divided into TFE group (150 mg·kg⁻¹·d⁻¹) and control group(received saline). HE staining was used to evaluate the morphologic difference of primary callus during the bone callus healing between these two groups. The relative expression of Runt-related transcription factor 2(Runx2) mRNA in the callus was identified by real-time polymerase chain reaction. Immunohistochemical technique was used to observe the Casein kinase 2-interacting protein 1(CKIP-1) protein level in the callus of the two groups. Maximum fracture load was tested by three point bend test.</p><p><b>RESULTS</b>The BMD, primary callus volume, trabecular member(Tb.N) and trabecular thickness(Tb.Th) were higher in TFE group than that in control group(<0.001). The Tb.N and Tb.Th of primary callus were higher in TFE group than control group (=0.001). The volume and bone volume/tissue volume of primary callus were in TFE group than control group(<0.01). The trabecular separation(Tb.Sp) of primary callus were in control group higher than TFE group(<0.01). The HE staining of the 6 week slices showed that the degree of cartilage ossification in callus of the TFE group was significantly higher than that in control group under high magnification. Real-time PCR revealed that the comparative expression of Runx2 mRNA in control group was higher than that in TFE group(<0.001); the positive number of CKIP-1 was less in TFE group than that in control group (<0.001). TFE could increase the maximum load of the primary callus (<0.001).</p><p><b>CONCLUSIONS</b>TFE can promote the cartiage ossification of callus in ovariectomized rats, enhancing the bone strength and bone quality in the process of fracture healing via the CKIP-1/Runx2 pathway.</p>

Identification of human serum protein targets of Qianggu Decoction () in primary type I osteoporosis based on tandem mass tag labeling and liquid chromatography-tandem mass spectrometry technology / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the serum protein targets of Qianggu Decoction (, QGD) on treating osteoporosis by the proteomics analysis using tandem mass tag (TMT) and liquid chromatographytandem mass spectrometry (LC-MS/MS).</p><p><b>METHODS</b>Twenty serum protein samples were recruited (10 patients with primary type I osteoporosis before and after QGD treatment) and the high abundance ratios protein was removed, two serum samples were extracted and labeled with TMT reagent. Then, mass spectrometric detection, identification of differentially expressed proteins and bioinformatics analysis of differentially expressed proteins were carried out.</p><p><b>RESULTS</b>A total of 60 proteins were identified, within a 99% confidence interval, to be differentially regulated of which, 34 proteins were up-regulated and 26 proteins were down-regulated. Differentially expressed proteins analyzed by Gene Ontology (GO) annotation mainly get involved in 12 different biological processes, 7 types of cellular components, and 6 kinds of molecular functions. Angiotensinogen (AGT), stromelysin-1 (MMP3), heparanase (HPSE) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were screened as candidate protein targets of QGD treatment, which were related to metabolic mechanism of bone remodeling and/or bone collagen of osteoporosis. By the utilization of the protein-protein interaction network analysis tool named STRING10.0, it showed that AGT, MMP3, HPSE and GAPDH were located in the key node of the protein-protein interactions network. Furthermore, AGT, MMP3, HPSE and GAPDH were found to be directly related to BMP, MAPK, Wnt, SMAD and tumor necrosis factor ligand superfamily member 11 (TNFSF11) families.</p><p><b>CONCLUSIONS</b>The proteomics analysis by using TMT combined with LC-MS/MS was a feasible method for screening the potential therapeutic targets associated with QGD treatment. It suggests that AGT, MMP3, HPSE and GAPDH may be candidate protein targets of QGD treatment which can be used as therapeutic effect monitor and early diagnosis of primary type I osteoporosis.</p>

The influences of DAO and SNGF in the treatment of hypothermia for neonates after birth asphyxia / 中国基层医药

Objective To study the changes of diamine oxidase (DAO)levels in neonates with hypoxic -is-chemic encephalopathy (HIE)treated with selective brain hypothermi,also to further evaluate Scores for Newborn Gastrointestinal Functior (SNGF).Methods 60 newborns with moderate and severe HIE who were in the NICU of our department from June 2013 to December 2014 were collected.The 60 newborns were randomly divided into hypo-thermia treatment group (HG)and conventional treatment group(CG).The serum was collected and ELISA method was used to test the consistency of DAO on admission and 7 days afterwards,respectively.Meanwhile,the SNGF level of the two groups at the two time points was compared.Results Neither the DAO and SNGF level of the two groups had statistical difference on admission(all P >0.05).Seven days later,both the DAO of the two groups and the SNGF decreased[(12.51 ±1.53)u/mL vs (7.88 ±1.87)u/mL,however,the variation of the hypothermia treatment group was apparently more significant than the change of the conventional treatment group(P =0.011).The SNGF scores of the two groups all decreased,while the hypothermia treatment group was significantly lower than the conventional treat-ment group,the difference were statistically significant (P =0.044,0.006,0.013).Besides,there was remarkably positive correlation between serum DAO level and SNGF (r1 =-0.825,r2 =-0.876,all P <0.05).Conclusion Hypothermia treatment could effectively reduce the injury of asphyxia neonatal gastrointestinal tract by inhibiting the level of DAO,thus improve the gastrointestinal function.

The changes and significance of diamine oxidase and intestinal fatty acid binding protein in the treat-ment of hypothermia for neonates after birth asphyxia / 中国小儿急救医学

Objective To study the changes of diamine oxidase (DAO )and intestinal fatty acid binding protein (I-FABP)levels in neonates with hypoxic-ischemic encephalopathy treated with selective brain hypothermia.Methods Collect a sample of 60newborns with moderate and severe hypoxic-ischemic encephalopathy who were hospitalized in the NICU of Matemal and Child Health Care Hospital of Baoding from June 2013to December 2014.The 60newborns were divided into two groups randomly:hypothermia group(n=30)and conventional treatment group(n=30).Selected 30cases hospitalized at the same period, except the related to the ischemia hypoxia and gastrointestinal dysfunction disease as the control group.The levels of serum levels of DAO and I-FABP were measured by ELISA on admission and 7days after treat-ment,respectively.And the score of gastrointestinal dysfunction were compared.Results Neither the levels of DAO and I-FABP in hypothermia group and conventional treatment group had statistical differences on ad-mission[DAO:(15.77±2.04)U/ml,(15.81±1.85)U/ml,P﹥0.05;I-FABP:(310.01±46.43)ng/L, (301.12±38.61)ng/L,P﹥0.05],but were higher than that in the control group [(7.65±0.74)U/ml, (51.65±6.91)ng/L].Seven days after treatment,both the levels of DAO and I-FABP of hypothermia group and conventional treatment group decreased [DAO:(7.88±1.87)U/ml,(12.51±1.53)U/ml;I-FABP:(59.16±6.17)ng/L,(121.31±21.54)ng/L],meanwhile,the variation of hypothermia group was more significant(P﹤0.05).The correlation of the plasma DAO and I-FABP levels and the score of gas-trointestinal dysfunction was significantly (r1=0.831,r2=0.827,P ﹤0.01).Conclusion Hypothermia treatment could effectively reduce the levels of DAO and I-FABP,thus improve the gastrointestinal function in some extent.

Clinical comparative study of vinorelbine and docetaxel separately combined with lobaplatin in treatment of recurrent and metastatic breast cancer / 中国基层医药

Objective To investigate clinical effect and safety difference of vinorelbine and docetaxel sepa-rately combined with lobaplatin in treatment of recurrent and metastatic breast cancer.Methods 160 patients with recurrent and metastatic breast cancer were chosen from July 2009 to July 2012 in our hospital,and they were random-ly divided into two groups,including A group (80 patients)with vinorelbine and B group (80 patients)with docetaxel on the basis of lobaplatin.The clinical efficacy for short -term,survival rate with follow -up and side effects incidence of both groups were compared.Results The RR and DCR of A group were 51.25%,68.75%,which of B group were 55.00%,71.25%.There was no significant difference in the clinical efficacy for short -term between the two groups (χ2 =1.04,2.37,all P >0.05).The survival rates in 1,2 and 3 years after treatment of A group were 60.00%,53.75%,28.75%,those of B group were 67.50%,57.50%,31.25%.There was no significant difference in the survival rate with follow -up between the two groups (χ2 =2.14,3.01,1.87,all P >0.05).The incidence rate of drug side effects of B group was significantly lower than A group(χ2 =13.14,9.33,15.74,11.65,8.29,all P <0.05).Conclusion Vinorelbine and docetaxel separately combined with lobaplatin in treatment of recurrent and metastatic breast cancer can efficiently control the disease progress and prolong the survival time;but docetaxel com-bined with lobaplatin treatment is helpful to reduce the risk of serious side effects.

Inhibition of Nonsmall Cell Lung Cancer Cell Migration by Protein Arginine Methyltransferase 1-small Hairpin RNA Through Inhibiting Epithelial-mesenchymal Transition, Extracellular Matrix Degradation, and Src Phosphorylation In Vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Protein arginine methyltransferases 1 (PRMT1) is over-expressed in a variety of cancers, including lung cancer, and is correlated with a poor prognosis of tumor development. This study aimed to investigate the role of PRMT1 in nonsmall cell lung cancer (NSCLC) migration in vitro.</p><p><b>METHODS</b>In this study, PRMT1 expression in the NSCLC cell line A549 was silenced using lentiviral vector-mediated short hairpin RNAs. Cell migration was measured using both scratch wound healing and transwell cell migration assays. The mRNA expression levels of matrix metalloproteinase 2 (MMP-2) and tissue inhibitor of metalloproteinase 1, 2 (TIMP1, 2) were measured using quantitative real-time reverse transcription-polymerase chain reaction. The expression levels of protein markers for epithelial-mesenchymal transition (EMT) (E-cadherin, N-cadherin), focal adhesion kinase (FAK), Src, AKT, and their corresponding phosphorylated states were detected by Western blot.</p><p><b>RESULTS</b>Cell migration was significantly inhibited in the PRMT1 silenced group compared to the control group. The mRNA expression of MMP-2 decreased while TIMP1 and TIMP2 increased significantly. E-cadherin mRNA expression also increased while N-cadherin decreased. Only phosphorylated Src levels decreased in the silenced group while FAK or AKT remained unchanged.</p><p><b>CONCLUSIONS</b>PRMT1-small hairpin RNA inhibits the migration abilities of NSCLC A549 cells by inhibiting EMT, extracellular matrix degradation, and Src phosphorylation in vitro.</p>

Seroprevalence of Toxoplasma gondii Among Primary School Children in Shandong Province, China

Although Toxoplasma gondii infection in primary school children has been investigated in many countries, limited surveys have been available in primary school children in China. In the present study, we report the seroprevalence of T. gondii infection in primary school children in Shandong province, China. Sera from 6,000 primary school children were evaluated for T. gondii antibodies with ELISA. The overall seroprevalence of T. gondii infection was 16.0% (961/6,000), of which 14.5% (870/6,000) were positive for anti-T. gondii IgG antibodies, 3.4% (206/6,000) positive for IgM, and 1.9% (115/6,000) were positive for both IgG and IgM. The results of the present investigation indicated a high seroprevalence of T. gondii infection in primary school children in Shandong province, China. Therefore, effective measures should be taken to prevent and control T. gondii infection in primary school children in this province. To the best of our knowledge, this is the first report of T. gondii seroprevalence in primary school children in Shandong province, China.

Inhibitory effect of Genipin on uncoupling protein-2 and energy metabolism of androgen-independent prostate cancer cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore whether the inhibitory effect of Genipin on uncoupling protein-2 (UCP-2) in mitochondria is involved in energy metabolism of androgen-independent PC3 prostate cancer cells.</p><p><b>METHODS</b>PC3 prostate cancer cells were cultured and treated with Genipin at the concentrations of 40, 80, and 160 μmol/L for 48 hours. Then the proliferation of the cells was detected by MTT assay, the expression of UCP-2 mRNA determined by RT-PCR, and the content of intracellular pyruvic acid (PA) and the activity of succinate dehydrogenase (SDH) in the mitochondria measured by visible spectrophotometry.</p><p><b>RESULTS</b>With the increased concentration of Genipin, the proliferative activity of the PC-3 cells, the expression level of UCP-2 mRNA, the content of intracellular PA and the activity of SDH in the cells were all decreased, namely, with the enhanced inhibitory effect of Genipin on UCP-2, a trend of reduction was observed in the proliferation of the cells, intracellular PA content, and SDH activity in the mitochondria.</p><p><b>CONCLUSION</b>Genipin is involved in the energy metabolism of androgen-independent PC3 prostate cancer cells by reducing the content of intracellular PA and the activity of SDH in the mitochondria, which may be associated with its inhibitory effect on UCP-2.</p>