AIM:To study the mechanism of Sihei-fang(SHF)in improving pigment deficiency disease(PD)by combining network pharmacology and me-tabolomics.METHODS:Using zebrafish embryos with pigment deficiency disease induced by 1-phe-nyl-2-thiourea(PTU)as an animal model,the ef-fects of SHF extract(0.01,0.02,0.04 mg/mL)on the morphology,melanin area,tyrosinase activity,and melanin content of zebrafish embryos were an-alyzed.Ultra high performance liquid chromatogra-phy-mass spectrometry(UHPLC-MS)was used to screen differential metabolites and obtain relevant metabolic pathways in the SHF treatment of mela-nin deficient zebrafish embryos model.Network pharmacology was used to obtain key targets for SHF treatment of PD and conduct KEGG pathway enrichment analysis.Import The identified differen-tial metabolites and SHF PD intersection targets were imported into the Metscape plugin,to estab-lish a"metabolite reaction enzyme gene"network,and search for key metabolites,targets,and meta-bolic pathways.RESULTS:SHF treatment could in-crease the formation of zebrafish melanin,activate tyrosinase activity,and increase melanin content.Metabolomics analysis obtained 54 differential me-tabolites,and metabolic pathway analysis was con-ducted on these metabolites,involving the biosyn-thesis of phenylalanine,tyrosine,and tryptophan,glycerol phospholipid metabolism,tyrosine metab-olism,linoleic acid metabolism,and aminoacyl tRNA biosynthesis pathways.Network pharmacolo-gy had obtained 55 cross targets of components and diseases.KEGG involved pancreatic cancer,TNF,cancer and other signal pathways.The joint analysis of metabolomics and network pharmacolo-gy identified four key targets:COMT,CYP1B1,TYR,and ALDH2;three key metabolites:L-tyrosine,ho-movanllate,L-lysine;three important metabolic pathways:tyrosine metabolism,valine/leucine/iso-leucine degradation,and lysine metabolism.CON-CLUSION:SHF has a good improvement effect on PD,and combined with metabolomics and network pharmacology,SHF may enhance its influence on the tyrosine metabolism pathway by regulating the metabolite L-tyrosine,thereby promoting the for-mation of melanin.

ObjectiveTo investigate the Alzheimer-associated neurofilament protein （AD7c-NTP） in urine of middle-aged and elderly people and its correlation between common metabolites. MethodsA total of 1 150 middle-aged and elderly people who did their physical exmanination in the health examination center of the Sichuan Science City Hospital and the Third Hopital of Mianyang were recruited from March 2017 to March 2020. The level of urine AD7c-NTP were measured by enzyme-linked immunosorbent assay （ELISA）， and common metabolites in blood were measured by biochemical analyzer. Based on urine AD7c-NTP level ≤1.5 ng/mL， the objects was divided into normal group （n=956） and elevated group （n=194）. Thier demographic data and blood biochemical indicators were collected. ResultsThe urine AD7c-NTP level in middle-aged and elderly people was 0.60（0.30~1.20） ng/mL. The urine AD7c-NTP level was higher in women than that in men ［1.04（0.40~1.30） ng/mL vs. 0.84（0.30~1.00） ng/mL， Z=4.202， P˂0.01］. And the urine AD7c-NTP level was lower in the normal group than that in the elevated group ［0.50（0.30~0.90） ng/mL vs. 2.10（1.70~2.10） ng/mL， Z=22.035， P˂0.01］. The results of the univariate comparison showed that， the differences between the two groups in age （Z=6.545）， fasting glucose （Z=3.506）， blood uric acid （Z=2.574）， urea nitrogen （Z=2.891）， creatinine （Z=2.243）， total bilirubin （Z=3.936）， glutathione （Z=0.969）， total cholesterol （t=3.956） and low density lipoprotein （Z=-5.678） were were statistically significant （P˂0.05 or 0.01）. Spearman correlation analysis showed that， the urine AD7c-NTP level was positively correlated with age and the levels of urea nitrogen， glucose， total cholesterol and low density lipoprotein （r=0.177， 0.178， 0.171， 0.109， 0.149， P˂0.01）， and negatively correlated with the level of total bilirubin （r=-0.172， P˂0.01）. Conclusionthe urine AD7c-NTP level in middle-aged and elderly females was signifitcantly higher than in middle-aged and elderly males.The urine AD7c-NTP level of middle-aged and elderly people was positively correlated with age， urea nitrogen， glucose， total cholesterol and low density lipoprotein， and negatively correlated with total bilirubin.

AIM: To study the effect of Qingdaipowder Gel (QDPG) on mice specific dermatitis (AD) model and the antibacterial effect of the ethanol extract of Qingdaipowder. METHODS: AD model of mice was established by repeated skin induction with 2,4-dinitrochlorobenzene (DNCB). Fifty-six mice were randomly divided into blank group, model group, Hydrocortisone Butyrate Cream group (Hyd, 1.5 mg/cm

Chinese hamster ovary (CHO) cells are the preferred host cells for the production of complex recombinant therapeutic proteins. Adenine phosphoribosyltransferase (APRT) is a key enzyme in the purine biosynthesis step that catalyzes the condensation of adenine with phosphoribosylate to form adenosine phosphate AMP. In this study, the gene editing technique was used to knock out the aprt gene in CHO cells. Subsequently, the biological properties of APRT-KO CHO cell lines were investigated. A control vector expressed an enhanced green fluorescent protein (EGFP) and an attenuation vector (containing an aprt-attenuated expression cassette and EGFP) were constructed and transfected into APRT-deficient and wild-type CHO cells, respectively. The stable transfected cell pools were subcultured for 60 generations and the mean fluorescence intensity of EGFP in the recombinant CHO cells was detected by flow cytometry to analyze the EGFP expression stability. PCR amplification and sequencing showed that the aprt gene in CHO cell was successfully knocked out. The obtained APRT-deficient CHO cell line had no significant difference from the wild-type CHO cells in terms of cell morphology, growth, proliferation, and doubling time. The transient expression results indicated that compared with the wild-type CHO cells, the expression of EGFP in the APRT-deficient CHO cells transfected with the control vector and the attenuation vector increased by 42%±6% and 56%±9%, respectively. Especially, the EGFP expression levels in APRT-deficient cells transfected with the attenuation vector were significantly higher than those in wild-type CHO cells (P < 0.05). The findings suggest that the APRT-deficient CHO cell line can significantly improve the long-term expression stability of recombinant proteins. This may provide an effective cell engineering strategy for establishing an efficient and stable CHO cell expression system.
Adenine/metabolism* ; Adenine Nucleotides ; Adenine Phosphoribosyltransferase/genetics* ; Adenosine Monophosphate ; Animals ; CHO Cells ; Cricetinae ; Cricetulus ; Recombinant Proteins/genetics*

Objective:To explore the effects of early warning nursing on preventing the incidence of venous thrombosis in patients with breast cancer after chemotherapy.Methods:Totally 103 patients with breast cancer admitted at the First Affiliated Hospital of Zhengzhou University from January to December 2018 who received routine nursing were included into the routine group, and 103 patients with breast cancer admitted at the First Affiliated Hospital of Zhengzhou University from January to December 2019 who underwent early warning nursing were included into the early warning group. The incidence of venous thrombosis 1 month after chemotherapy was compared between the two groups. Color Doppler ultrasonography was used to detect axillary blood flow before and 1 month after chemotherapy.Results:The incidence of venous thrombosis in the early warning group after chemotherapy was lower than that in the routine group, and the difference was statistically significant ( P<0.05) . The maximum blood flow rate and average blood flow rate in the armpit after chemotherapy in the early warning group were higher than those in the routine group, with statistically significant difference ( P<0.05) . Conclusions:Early warning nursing has a positive effect on the incidence of venous thrombosis in patients with breast cancer after chemotherapy, which can improve their hemagglutination.

Background Tumstatin is the most active endogenous angiogenesis inhibitor,which has a marked inhibitory effect on pathological neovascularization,and Tum5 is an angiogenesis inhibitors fragment of fulllength tumstatin.Objective This study was to investigate the effects of adenovirus-mediated overexpression of recombinant Tum5 gene on the proliferation,migration and tube formation of human umbilical vein endothelial cells (HUVECs) in physiological status.Methods The empty adenoviral vector expressing green fluorescent protein (rAd-GFP) and the viral vector expressing recombinant Tum5 gene were constructed.The HUVECs cultured in RPMI1640 medium were divided into normal control group,empty vector group (rAd-GFP group) and Tum5 gene infection group (rAd-GFP-Tum5 group).The rAd-GFP and rAd-GFP-Tum5 adenoviral particles at the density of 1 × 1010/ml were added into the medium to infect the cells for 48 hours.The proliferation of the cells was assayed at 24,48 and 72 hours by cell counting kit-8 (CCK-8) to evaluate the proliferative rate;the migration number of the cells was detected at 48 hours after infection by Transwell chamber;the tube formation number of the cells were detected by Matrigel method.The concentration of vascular endothelial growth factor (VEGF) in cell supernatants was assayed by ELISA at 24,48,and 72 hours following adenoviral infection.Results The cultured cells showed green fluorescence in the rAd-GFP group and rAd-GFP-Tum5 group under the inverted fluorescence microscope,and the infection efficiency of rAd-GFP and rAd-GFP-Tum5 was 55.13％ and 50.31％,respectively.No significant difference was found in cell proliferative rate among normal control group,rAd-GFP group and rAd-GFP-Tum5 group both at 24 and 48 hours after infection (both at P＞0.05),and the cell proliferative rate was significantly lower in the rAd-GFP-Tum5 group than that in the normal control group and rAd-GFP group at 72 hours after infection (both at P＜0.01).The migration number of the cells at 48 hours after infection was 2 260.25-±930.44,2 370.00±441.06 and 723.75± 363.80 in the normal control group,rAd-GFP group and rAd-GFP-Tum5 group,showing a significant difference among the groups (F =8.524,P =0.008),and the migrated cells were evidently decreased in the rAd-GFP-Tum5 group compared with the rAd-GFP group and the normal control group (both at P＜ 0.01).The tube number at 48 hours after infection was 95.67±5.86,88.00±4.58 and 20.67±3.51 in the normal control group,rAd-GFP group and rAdGFP-Tum5 group,showing a significant difference among the groups (F=226.498,P＜0.01),and the tube number in the rAd-GFP-Tum5 group was significantly reduced in comparison with the normal control group and rAd-GFP group (both at P＜ 0.01).The considerably differences in VEGF concentration in the cell supernatants were found in different groups and various time points (Fgroup =73.260,P＜0.01;Ftime =73.477,P＜0.01),and VEGF concentration in the cell supernatants was significantly decreased in the rAd-GFP-Tum5 group compared with the rAd-GFP group at both 48 hours and 72 hours (both at P＜0.01).Conclusions The overexpression of the recombinant Tum5 can inhibit the proliferation,migration and tube formation of the HUVECs in physiological status,which may be associated with Tum-5-mediated down-regulation of VEGF protein in the cell supernatant.

Objective To explore the impact of prenatal diagnosis on the early treatment and short and medium term outcome of neonatal pulmonary atresia with intact ventricle septum (PA/IVS) or critical pulmonary stenosis with intact ventricle septum (CPS/IVS). Methods According to the case-control method, twenty-eight neonates with (PA/IVS) or (CPS/IVS) who had percutaneous pulmonary balloon valvuloplasty (PBPV) surgery indications, were divided into the prenatal diagnosis group (n?=?15) and the postnatal diagnosis group (n?=?13). The prenatal diagnosis group was diagnosed in fetal period and the intervention program was since developed . The postnatal diagnosis group was referred from other hospitals, and the intervention program was developed after conifrmation of the diagnosis. All the neonates accepted the PBPV surgery after the hemodynamic parameters stable. All neonates were followed-up at 1 month, 3 months, 6 months, 1 year, and 2 years after surgery. Clinical situations, echocardiography results, and interventional cardiology measurements were compared between two groups. Result The average age and weigh was 7.53?±?3.18 days and 3102.32?±?708.40 g respectively at the time of PBPV surgery in 28 neonates. Among them, 9 neonates were PA/IVS and 19 neonates were CPS/IVS. The mean follow-up time was 18.8?±?5.22 months and there were no death. The ages at admission and at the ifrst treatment were signiifcantly younger in the prenatal diagnosis group than those in the postnatal diagnosis group (P??0.05). Conclusion Prenatal diagnosis is helpful for the early intervention in neonates with PA/IVS and CPS/IVS, and can reduce the complications after surgery.

The Strem-1 is a member of the immunoglobulin family,which is recently found to be closely associated with the inflammatory and a sensitive marker of the inflammatory response.Many pathogenic microorganisms infection can make sTREM-1 highly expressed,it is involved in the secretion of pro-inflammatory factors by the role of TOLL receptors,and played an important role in the development of sepsis.It is a more sensitive and reliable indicator in the diagnosis and monitoring of sepsis in recent years.It is necessary to study the characteristics and role of sTREM-1 in the development of sepsis,and it has important significance in preventing the occurrence of sepsis and reasonable treatment or prognosis evaluation.

OBJECTIVETo establish a HPLC-ELSD method for simultaneous determination of trillin and desgalactotigonin contents in Solanum lyratum.

METHODA Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) was adopted, with the mobile phase consisting of acetonitrile-10 mmol x L(-1) ammonium acetate (52: 48). The temperature was 25 degrees C, the flow rate was set at 0.6 mL x min(-1), and the sample size is 20 microL. The temperature of drift tubes and gas flow rate of the detector were set at 95 degrees C and 2.3 L x min(-1), respectively.

RESULTWith in the linear ranges of 20-200 mg x L(-1) and 10-100 mg x L(-1), trillin and desgalactotigonin show a good linear relationship. The average recovery was 99.4% (RSD 0.90%) for trillin and 100.3% (RSD 1.1%) for desgalactotigonin.

CONCLUSIONThe method is so accurate and easily reproducible that it is suitable for the quality control of S. lyratum medicinal materials.

Chromatography, High Pressure Liquid ; Drug Stability ; Drugs, Chinese Herbal ; analysis ; chemistry ; Quality Control ; Reproducibility of Results ; Scattering, Radiation ; Solanum ; chemistry

Objective To investigate the association between matrix metalloproteinase (MMP) -2 C735T and MMP-9 C1562T polymorphisms and TOAST subtypes, the outcome in patients with stroke. Methods A total of 232 patients with ischemic stroke were divided into large artery atherosclerosis (LAA, n =37), cardioembolism (CE, n =31), small artery occlusion (SAO, n =65) stroke, stroke of other demonstrated etiology (SOE, n =2), and stroke of undemonstrated etiology (SUE, n =97) according to TOAST criteria. A total of 235 healthy subjects in the outpatient served as control. Genetic polymorphisms of MMP-2 C735T and MMP-9 C1562T were identified by polymerase chain reaction-restriction fragment length polymorphism.The outcome of patients was evaluated with Barthel Index (BI) at day 21 and 90 after stroke.Results The frequencies of MMp-2 735CC genotype and C allele in the ischemic stroke group (CC genotype: 63.36％ vs. 54.04％,x2 =4. 182, P=0.014; C allele: 79.31％vs. 74.04％,x2 =3. 936, P =0. 047 ) and its LAA subtype ( CC genotype: 78. 37％ vs. 54. 04％, x2 =7. 740, P =0. 005; C allele: 87. 83％ vs. 74. 04％, x2 =6. 655, P =0. 01 ) were significantly higher than those in the control group. The frequencies of MMP-9 1562CT +TT genotype and T allele in the ischemic stroke group (CT +TT genotypes: 21.98％ vs. 13. 19％,x2 =6. 233, P=0.013; T allele: 11.64％ vs. 7. 02％ ,x2 =5. 891, P =0. 015)and its LAA subtype(CT +TT genotypes: 32. 43％ vs. 13. 19％ ,x2 =8. 892, P =0. 003; T allele: 20. 27％ vs. 13.19％ ,x2 =13. 950, P =0. 000). Multivariate logistic regression analysis indicated that risk of ischemic stroke and its LAA subtype with MMP-2 735CC genotype (ischemic stroke: odds ratio [OR]1.099, 95％ confidence interval [CI] 1.038-1.260, P =0.028; LAA: OR 1.360, 95％ CI 1. 167-5. 774, P =0. 009) and with MMP-9 1562TT genotype (ischemic stroke: OR 9. 409,95％ CI 1. 154-76. 722, P =0. 036; LAA: OR 8. 962, 95％ CI 1. 380-58. 218, P =0. 022)increased significantly. There were no significant correlation between the different genotypes of MMP-2 and MMP-9 and the outcome of ischemic stroke. Conclusions Polymorphisms of MMP-2 C735T and -9 C1562Tare associated with ischemic stroke and its subtype large artery atherosclerotic stroke, but not associated with the outcome in patients with ischemic stroke