Serological characteristics of anti-PP1Pk and literature review on P blood group system / 中国输血杂志

【Objective】 To comprehensively explore the serological characteristics of anti-PP1Pk and potential therapeutic strategies for recurrent miscarriage in p-blood type pregnant women. 【Methods】 Neutralization with pigeon egg white and human plasma, disruption of IgM antibodies by 2-mercaptoethanol reagent, and complement adding were conducted. Anti-PP1Pk titers under different processing conditions, media and temperatures were determined, and neutralizing effect of human plasma on anti-PP1Pk and its sensitization complement ability were analyzed. 【Results】 The titers of anti-PP1Pk in saline and column agglutination were 4 and 8, respectively. Low temperature increased titers, while β-mercaptoethanol treatment significantly reduced them. Pigeon egg white partially neutralized anti-PP1Pk antibodies. Human plasma was also capable of reducing anti-PP1Pk titers with neutralization capability surpassing that of pigeon egg white, and there were individual differences in neutralization capability. 【Conclusion】 The anti-PP1Pk was a blend of antibodies encompassing both IgM and IgG types, exhibiting cold reactivity, and having the potential for complement activation. Human plasma emerges as an effective modulator for diminishing the efficacy of anti-PP1Pk. Plasma transfusion holds promise as a therapeutic avenue for addressing recurrent miscarriages in pregnant individuals with the p phenotype.

Effect of printing orientation on physical and mechanical properties of 3D printing prosthodontic base resin materials / 北京大学学报(医学版)

Objective:To analyze the influence of forming direction on the surface characteristics,elastic modulus,bending strength and fracture toughness of printed parts and the relationship between forming direction and force direction,and to provide scientific basis and guidance for the clinical applica-tion of oral denture base resin materials.Methods:The 3D printing technology was used to print denture base resin samples.The shape and size of the samples referred to the current standard for testing conven-tional denture base materials.The samples used for physical performance testing were cylindrical(with a diameter of 15 mm and a thickness of 1 mm)and printed at different angles along the Z axis(0°,45°,90°).Scanning electron microscope was used to observe the microscopic topography of the different sam-ples.The color stability of different samples was observed by color stabilizer.The surface roughness of the samples was analyzed by using surface roughness tester.The Vickers hardness was measured to ana-lyze the hardness of the samples.The samples used for mechanical performance testing were rectangular(elastic modulus and bending strength:A length of 64 mm,a width of 10 mm,and a height of 3.3 mm;fracture toughness:A length of 39 mm,a width of 8 mm,and a height of 4 mm),divided into two groups:W group and H group.The W group was printed from the bottom up along the Z axis with the length × width as the bottom surface parallel to the X,Y axis plane,while the H group printed from the bottom up along the Z axis with the length × height as the bottom surface parallel to the X,Y axis plane.The forming angles of both groups were equally divided into 0°,45°,and 90°.The elastic modulus,bending strength and fracture toughness of different samples were studied through universal mechanical testing machine.SPSS 22.0 software was used for statistical analysis.Results:The microscopic topogra-phy and roughness of different samples were closely related to the printing direction,with significant differences between the 0°,45°,and 90° specimens.The 0° specimens had the smoothest surface(roughness＜1 μm).The surface of the 45 ° specimen was the roughest(roughness＞3 μm).The microhardness of the 0° sample was the best[(196.13±0.20)MPa],with a significant difference com-pared with the 90° sample[(186.62±4.81)MPa,P＜0.05].The mechanical properties of different samples were also closely related to the printing direction.The elastic modulus,bending strength,and fracture toughness of the 45° samples in the W group were the highest compared with the other groups.The results of elastic modulus showed that in the H group,the 45° specimens had the highest elastic mo-dulus,which was significantly different from the 0° and 90° specimens(P＜0.05).The elastic modulus of 0° and 45° specimens in the W group were higher than those in 90° specimens(P＜0.05).The bending strength results showed that there was no significant difference between the specimens from dif-ferent angles in the H group.The bending strength of the 90° specimens in the W group was the smallest,and there was a significant difference between 90° and the 0° and 45° specimens(P＜0.05);And the bendind strength of the 0° and 45° specimens in the W group was significantly higher than that of the 0° and 45° specimens in the H group(P＜0.05).The fracture toughness results showed that the fracture toughness of the H group specimens was lower than 1.9 MPa m1/2,which was specified in the denture base standard.The 45° samples in the W group were the highest,with significant differences compared with the 0° and 90° samples(P＜0.05).And the 90° samples of the W group specimens were lower than 1.9 MPa m1/2.And the fracture toughness of the 45° specimen in the W group was significantly higher than that of all the specimens in the H group(P＜0.05).Conclusion:The 0° samples had rela-tively better physical properties.The 45° samples had the best mechanical properties.But the fracture toughness of specimens(H group and 90° samples of W group)did not yet meet clinical requirements.That indicated that the characteristics of the 3D printing denture base resin were affected by the printing direction.Only when the performance of the printed samples in all directions met the minimum require-ments of the standard,they could be used in clinical practice.

Frequency survey of blood group PP1PK in Chinese Lahu Population / 中国输血杂志

【Objective】 To investigate the frequency of P1PK and GLOB blood group in Chinese Lahu population and their genetic status, so as to provide data support for the safety of blood transfusion and give advice and transfusion guidance for pregnant women. 【Methods】 Unrelated individuals of Chinese Lahu population were randomly selected for serological identification of P1PK and P blood group and gene sequencing analysis. The frequency of P1PK and GLOB blood group were analyzed. 【Results】 Six cases of anti-PP 1Pk(formerly known as anti-Tja) negative blood type were identified as the rare P1-PK-P- blood type (formerly known as Tja- blood type or p blood type, hereinafter referred to as p blood group) from 300 Lahu population, with phenotypic frequency of p blood group in P1PK and GLOB blood group system in Lahu population at 2.0%(6/300). 【Conclusion】 The phenotypic frequency of blood group p in Lahu population was significantly higher than that in Europe (5.8 persons per million) and Hong Kong, China (1 person per million),indicating significant ethnic specificity and regional ethnic differences.

Simultaneous determination of lacosamide and perampanel concentration in human plasma by LC-MS/MS / 中国药房

OBJECTIVE To establish a method for simultaneous determination of two third-generation anti-epileptic medicines such as lacosamide and perampanel in human plasma and apply this method in clinical practice. METHODS Using clozapine as internal standard， the concentrations of lacosamide and perampanel of plasma samples in 10 epileptic patients were determined by LC-MS/MS after protein precipitation with acetonitrile and dilution with acetonitrile-water （20∶80，V/V）， and the plasma minimum concentrations were obtained by dilution of multiple. The determination was performed on Welch Ultimate XB-C18 column， with mobile phase A consisted of 10 mmol/L ammonium formate and mobile phase B consisted of methanol-acetonitrile-isopropanol （0.2% formic acid） mixed solution （7∶1.5∶1.5， V/V/V） for gradient elution at the flow rate of 0.4 mL/min. The column temperature was set at 40 ℃ ， and the sample size was 5 μL. The electrospray ion source and multi-reaction monitoring mode were used for positive iron scanning. The ion pair used for quantitative analysis of lacosamide， perampanel and internal standard were m/z 251.2→ 144.1， m/z 350.2→219.2 and m/z 327.2→270.0， respectively. RESULTS The linear ranges of lacosamide and perampanel were 0.001 25-0.125 μg/mL（r＞0.99）， 0.037 5-3.75 ng/mL （r＞0.99）； the limits of quantification were 0.001 25 μg/mL and 0.037 5 ng/mL， respectively. The precision and accuracy within and between batches， extraction recovery rate， matrix effect， and stability all met relevant requirements. The minimum concentrations of lacosamide in No. 1-5 patients were 5.3-12.2 μg/mL， and the minimum concentrations of perampanel in No.6-10 patients were 208-510 ng/mL， respectively. CONCLUSIONS The established method is simple， rapid and suitable for the therapeutic drug monitoring of lacosamide and perampanel.

Analysis of alterations in early postoperative pulmonary bacterial microbiome characteristics in lung transplant recipients / 中华器官移植杂志

Objective:To explore the abundance, diversity, and structural changes of early postoperative pulmonary bacterial microbiota in lung transplant recipients.Methods:Recruiting 40 recipients who underwent lung transplantation surgery at the First Affiliated Hospital of Guangzhou Medical University from October 2020 to May 2022 for the study.All recipients did not receive antibiotic treatment within 4 weeks prior to surgery, and all recipients received a unified immunosuppressive and anti infection regimen after surgery.The bronchoalveolar lavage fluid(BALF) was collected from the amputated lung in vitro before the transplantation for 16S ribosomal RNA sequencing and flora analysis.BALF was also collected at the scheduled time from the transplanted lung on the 7th, 14th and 30th days post transplantaion for analysis.Results:The study included a total of 40 recipients who did not receive antibiotic treatment within 4 weeks before surgery, including 35 males.Among the study participants, there were 14 cases of primary obstructive pulmonary disease, 19 cases of interstitial lung disease, 3 cases of occupational lung disease, and 4 others.Microbiome in BALF of transplanted and detached autologous lungs at the first week after surgery α( P<0.05) and β diversity is statistically significant( R2=0.08, P=0.001), and the bacterial community in the transplanted lungs α Diversity is lower than that of explant lungs.Starting from the second week after surgery, the richness and species diversity of the transplanted lung microbiota gradually increase.The bacterial structure was also changed with postoperative time, and the relative abundance of the same bacterial species were varied at different time points.The bacterial community in BALF was mainly dominated by Proteobacteria both explant lungs and transplant lungs.The relative abundance of Staphylococcus and Acinetobacter genera at the BALF in transplanted lungs was higher than that in explant lung samples, but their relative abundance decreased over time after surgery. Conclusions:The α diversity of the early postoperative pulmonary microbiota after lung transplantation was lower than that of the amputated autologous lung, and the bacterial richness and species diversity in the microbiota of the transplanted lung gradually increased at the second week after the transplantation.The bacterial microbiota of the transplanted lung is changed complicatedly with time.

Overview of transfusion-related human red cell immunity / 中华微生物学和免疫学杂志

Red blood cells are the body′s most abundant cells, constantly providing oxygen and removing CO 2. Since the discovery of ABO blood group, blood transfusion technology, as an important safeguard means, has continuously promoted the development of modern medical technology. Currently, 45 erythrocyte blood group systems, more than 300 antigens, and many more subtypes and variants have been identified in humans. These antigens show great differences in the process of transfusion-related immunity, and some red blood cell antigens and their antibodies that have great influence on transfusion have also been studied in detail, such as antibodies to antigens in the Rh and Kidd systems. Even so, the hemolytic transfusion reaction caused by blood group antibodies is still difficult to avoid, and blood transfusion workers need to make continuous progress in transfusion strategies and detection techniques.

Distribution of ABO, Rh, Mn, P, h blood groups and gene frequency in Lahu ethnic minority in Yunnan / 中国输血杂志

【Objective】 To understand the distribution and gene frequency of main red blood cell blood groups in Lahu ethnic minority and analyze the genetic characteristics of Lahu people. 【Methods】 1) ABO forward and reverse typing had been performed by microplate method; 2) Rh, MN, H, P1Pk and Mur antigen were tested by the tube method. If the ABO forward and reverse typing were incompatible, the tube method was used for confirmation. 【Results】 The distribution characteristics of blood group and gene frequency in Lahu ethnic minority were as follows: B>O>A>AB for ABO, with genotype frequency as p 11.1%, q 27.5% and r 61.4%; the frequency of Rh genotype was CDe 83.3%, cDE 12.0%, cDe 2.42%, CDE 2.32%, CdE 0%, Cde 0%, cdE 0% and cde 0%; M > MN>N for MN blood group, with genotype frequency as M 75.26% and N 24.74%; P1

Application of MALDI-TOF MS in clinical difficult blood group typing / 中国输血杂志

【Objective】 To explore the application of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) in the genotyping of difficult blood typing samples, and to provide evidence for clinical blood transfusion. 【Methods】 Three ambiguous blood group samples, submitted to Shanghai Blood Center by Shanghai regional hospitals, were studied, of which Sample1 included the proband and his parents. Serological methods were used to perform blood group typing, direct antibody test, unexpected antibody screening and identification test. Blood group genotyping was performed by using the MALDI-TOF MS detection systeme stablished in our laboratory. Sanger sequencing was used to confirm gene mutation sites, and serological or flow methods were used to verify specific samples′ phenotype. 【Results】 Serological results indicated the existence of antibodies against high frequency antigens in sample 1 (including proband and her mother), 2 and 3. The genotyping results of MALDI-TOF MS showed that the proband of sample 1 was Di(a+ b+ ), her father was Di(a-b+ ), her mother was Di(a+ b-), sample 2 was p, and sample 3 was Jr(a-). Sequencing results of three samples were consistent with mass spectrometry typing results. Serological results showed that sample 2 had a p phenotype. The flow cytometry results suggested that sample 3 had a Jr(a-) phenotype. 【Conclusion】 For the first time, we applied MALDI-TOF MS technology to blood type genotyping of ambiguous clinical samples in China. Compared with other genotyping methods such as PCR-SSP, MALDI-TOF MS has the advantages of rapid detection, high throughput and high specificity, which would contribute to identification of difficult blood typing samples in the future, as well as rare blood group screening.

The clinical serological characterization of cisAB and B(A) blood groups / 中国输血杂志

【Objective】 To determine the rare ABO blood subgroups rapidly and ensure the blood transfusion safety of five patients by a series of serological tests and family investigation, as their preliminary serological results of ABO blood grouping was inconsistent. 【Methods】 ABO blood grouping, antibody screening and Coombs′ tests were performed by the routine serological methods, including manual tube and automatic blood group analyzer, which had matched micro-column gel cards from Diagnostic Grifols. Polymerase chain reaction (PCR) was used to amplify the 6 and 7 exons as well as their adjacent intron region of ABO gene. The patients and their relatives′ ABO blood group and subgroup were analyzed and identified through the comparison with serological phenotype database of ABO blood group. The products of PCR were sequenced directly, and the gene mutation was identified through the comparison with the Blood Group Antigen Gene Mutation Database. 【Results】 Whether micro-column gel cards or manual tube test, the forward and reverse tests of serological grouping were not supported by each other on the five patients′ ABO blood grouping. The forward tests of patients No.1~3 showed AwB phenotype and the reverse tests showed B group. No.4 patient was the forward ABw phenotype and the reverse A group, and No.5 patient was the forward normal AB phenotype and the reverse B group, respectively. All of 5 patients′ Rh C/D/E blood grouping showed clearly; the IDT test and antibody screening result of patient No.1 was positive, while the antibody screening result of patient No.4 was negative. 【Conclusion】 The blood group serological characterization of patient No.1~3 met B(A) blood group, and patient No.4~5 met CisAB blood group. These tests can make a preliminary diagnosis of blood group phenotype, which are verified correctly via blood group genotype.

Quantitative detection of red blood cell antibody-mediated complement activation / 中国输血杂志

【Objective】 To construct an in-vitro model of erythrocyte antibody-mediated complement activation, and establish quantitative detection methods based on flow cytometry and spectrophotometry, so as to explore the correlation of anti-body titers and complement activation speed, and provide a methodological basis for studying the adverse transfusion reactions of anti-body mediated complement hemolysis. 【Methods】 Mouse monoclonal antibody that recognized human C3b and fluorescent secondary antibody were used to label C3b fragments on erythrocytes, and the deposition of C3b fragments after complement activation was detected by flow cytometry. The absorbance at 540 nm of the supernatant in the complement activation reaction system was measured by spectrophotometry as the amount of hemoglobin released was related to the absorbance. 【Results】 The complement activation system was constructed according to the ratio of 3% red blood cell suspension (mixed for 6 people) 1∶anti-Tja 1∶complement 2. The repeatability was good (P value>0.05) as different red blood cell mixtures had been used to repeat the detection reaction system. When using 32×, 64× and 128× dilutions of anti-Tja mediated complement activation, the deposition of C3b fragments has been detected by flow cytometry at 30 s, 1 min and 2 min, respectively, and MFI peaked at 5 min, 10 min and 30 min, respectively. No obvious hemolysis has been observed within 1.5 h. 【Conclusion】 In vitro model of anti-Tja-mediated complement activation demonstrates the speed of complement activation is related to the concentration of antibody. At a certain antibody concentration, the speed of complement activation has been slowed down, and no obvious hemolysis observed.

Preparation of human monoclonal anti-C cell line from peripheral blood B lymphocytes of D--donor / 中国输血杂志

【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.

Baicalin protects against 17α-ethinylestradiol-induced cholestasis via Sirt1/HNF-1α/FXR pathway / 中国药理学与毒理学杂志

OBJECTIVE Baicalin is a major flavonoid component of Scutellaria baicalensis, and has been used in the treatment of liver diseases for many years. However, the role of baicalin in estrogen-induced cholestasis (EIC) remains to be elucidated. This present study explored the protective effect of baicalin against estrogen-induced liver injury and further elucidated the mechanisms involved both in vivo and in vitro. METHODS We conducted a series of experiments using 17α-ethinylestradiol (EE) induced cholestatic rats and cultured HepG2 cells. Serum, bile, and liver samples were collected for biochemical and histological analyses. Bile acid composition in liver was analyzed by LC-MS/MS. The mechanisms underlying the hepatoprotective of baicalin were investigated by RT-PCR, Western blotting analyses and immunohistochemistry. RESULTS Baicalin showed obvious hepatoprotective effects in EIC rats by reducing serum bio?markers and increasing the bile flow rate, as well as by alleviating liver histology and restoring the abnormal composition of hepatic bile acids (BAs). In addition, baicalin protected against EE induced liver injury by up-regulation of the expres?sion of hepatic efflux transporters and down-regulation of hepatic uptake transporters. Furthermore, baicalin increased the expression of hepatic BA synthase (CYP27A1) and metabolic enzymes (Bal, Baat and Sult2a1) in EIC rats. We showed that baicalin significantly inhibited hepatic inflammatory responses in EIC rats through reducing elevated levels of TNF-α, IL-1β, IL-6 and NF-κB. Finally, we confirmed that baicalin maintains BA homeostasis and alleviates inflamma?tion through Sirt1/HNF-1α/FXR signaling pathway. CONCLUSION Baicalin protects against estrogen-induced cholestatic liver injury, and the underlying mechanism involved is related to activation of the Sirt1/HNF-1α/FXR signaling pathway.

-D- phenotype screening by polybrene method / 中国输血杂志

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

Genetic polymorphism and antibody screening of RhD negative Chinese Han population in Jiayuguan / 中国输血杂志

【Objective】 To investigate and analyze the polymorphism of RHD gene in RhD-negative population in Jiayuguan using molecular biological technique, so as to accurately identify RhD-negative individuals, and formulate individualized transfusion strategies. 【Methods】 The RhD negative voluntary blood donors and patients (mainly pregnant women) were recruited. After informed consent, history of blood transfusion and pregnancy of them were investigated, and samples were collected for negative D confirmation, gene sequencing as well as antibody screening and identification. 【Results】 Among the 96 samples, 73 cases were RHD gene deletion, 18 RHD*01EL.01(17 RHD1227A homozygous type and 1 RHD1227A heterozygous type), 2 weak RHD*15 type (845G/A), 1 partial D type, i. e. RHD-CE(7) -D heterozygous allele, and 2 RHD*01N.16 variant. Antibody was detected out in 4 cases, among which 2 were positive for anti-D, 1 anti-D plus anti-E, and 1 anti-Dia. 【Conclusion】 The proportion of DEL gene in RhD negative Chinese Han population in Jiayuguan is slightly lower than that in general Chinese Han population. No anti-D or RHD-HDN was observed in DEL type women due to multiple pregnancy or delivery of D positive newborns.

Interference of CD38 monoclonal antibody in blood compatibility testing and its countermeasures: A general consensus among experts / 中国输血杂志

With continuous discovery of tumor immune targets and continuous changes in antibody research and development technology, antibody drugs are becoming more and more widely used in clinical practice. However, some targets are not only expressed on tumor cells, but also on red blood cells. Therefore, the clinical application of antibodies against the corresponding targets may interfere with the detection of blood transfusion compatibility, resulting in difficulty in blood matching or delay of blood transfusion. This consensus summarizes the current solutions for the interference of CD38 monoclonal antibody (CD38 mAb) in transfusion compatibility testing. After analyzing the advantages and disadvantages of different methods, polybrene and sulfhydryl reducing agents [dithiothreitol (DTT) or 2-mercaptoethanol (2-Me)], as a solution for CD38 mAb interference in blood compatibility testing, are recommended for Chinese patients, so as to eliminate blood transfusion interference produce by CD38 mAb and further provide a pre-transfusion workflow for clinicians and technicians in Department of Blood Transfusion.

Changes in the proliferation and angiogenesis of human dental pulp stem cells after treated with prostaglandin E1 combined with basic fibroblast growth factor / 中国组织工程研究

BACKGROUND: Prostaglandin E1 and basic fibroblast growth factor can promote the proliferation of human dental pulp stem cells, but the effects of their combinations on the proliferation of human dental pulp stem cells and angiogenesis are unknown. OBJECTIVE: To investigate the effects of prostaglandin E1 combined with basic fibroblast growth factor on the proliferation and angiogenesis of human dental pulp stem cells. METHODS: (1) Human dental pulp stem cells were isolated and cultured in vitro. After detection and identification of surface markers, prostaglandin E1 and basic fibroblast growth factor at concentrations of 5, 10, 20, 50 and 100 μg/L were used to treat human dental pulp stem cells in vitro. The untreated cells served as control group. The cell proliferation was detected by cell counting kit-8 assay, and the optimum drug concentration and time of drug action were screened. (2) The in vitro cultured human dental pulp stem cells were divided into four groups: blank control group, prostaglandin E1 group, basic fibroblast growth factor group and combination group. The cell proliferation was detected by cell counting kit-8 assay. Human dental pulp stem cell conditioned medium was extracted. The levels of vascular endothelial growth factor and endostatin in the culture medium were detected by ELISA. The in vitro tubular formation ability of human umbilical vein endothelial cells after culture in conditioned medium was tested by tubule formation experiment. RESULTS AND CONCLUSION: The optimum concentration of prostaglandin E1 and basic fibroblast growth factor was 20 μg/L, and the optimum time of action was 2 days. Compared with the blank control group, the relative proliferation rate, level of vascular endothelial growth factor and the angiogenesis ability of human umbilical vein endothelial cell in vitro in the prostaglandin E1, basic fibroblast growth factor and combination groups were significantly increased (P < 0.05), while the level of endostatin was significantly decreased (P < 0.05). All above index levels in the combination group were significantly superior to those in the prostaglandin E1 and basic fibroblast growth factor groups (P < 0.05). In summary, prostaglandin E1 combined with basic fibroblast growth factor can promote the proliferation of human dental pulp stem cells and enhance the tubular formation ability of human umbilical vein endothelial cells in vitro.

Mechanism of Calculus Bovis Sativus in inhibiting hepatocyte lipid deposition based on serum pharmacology / 中国中药杂志

The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.

Effect and Mechanism of Pilose Antler Different Components on Bone Tissue of Ovariectomized Osteoporosis Model Rats / 中国实验方剂学杂志

Objective:To explore the effect and mechanism of pilose antler different components on the bone tissue of ovariectomized osteoporosis model rats and ascertain the material basis of pilose antler. Method:fifty-six SD rats were divided randomly into seven groups:normal group,model group,Xianling Gubao group(468 mg·kg-1),Bujiale group(80 mg·kg-1),polysaccharide group(50 mg·kg-1),polypeptides group(175 mg·kg-1),polysaccharide and polypeptide mixture group(50 mg·kg-1+175 mg·kg-1). Osteoporosis mode was established through ovary resection of female rats,meanwhile,the rats were given different components of pilose antler for consecutively 12 weeks. Subsequently, using absorptiometry to measure the rats' bone mass density. The activities of bone alkaline phosphatase(BALP),osteocalcin (OT),bone morphogenetic protein2(BMP-2),Smad1,Smad5,Runt-related transcription factor 2 (RUNX2) were detected by enzyme-linked immuno sorbent assay (ELISA). The expression of BMP-2,Smad1,Smad5,Runx2 protein was examined by Western blot and Real-time polymerase chain reaction (Real-time PCR). Morphological assay for bone tissue were detected by htoxylin eosin(HE) staining. Result:After 12 weeks, Compared with the normal group, the osteoporosis model group showed significantly decrease in bone mineral density(PPPConclusion:Pilose antler different components has therapeutic effect on ovariectomized osteoporosis model rats.The mechanism may be related to up-regulat the expression of BMP-2/Smad1,Smad5/Runx2 signal pathways.

Modern Research of Calculus Bovis ( First):Retrospect and Prospect / 医药导报

Calculus Bovis is a valuable traditional Chinese medicine and has been used for more than two thousand years in clinic with the effects of puring heart, sweeping phlegm, resuscitation, extinguishing wind and detoxification. Since the founding of the People's Republic of China, modern methods have been utilized by traditional Chinese medicine researchers in the resource identification, chemical components, pharmacodynamics, pharmacokinetics, pharmacy, clinical application, etc. It is their continually exploration that makes significant achievements for the modern research of Calculus Bovis. This article statistically analysed the literatures from 1949 to December 2015 in Pubmed, CNKI, Wanfang, Vip database etc. to review Calculus Bovis and its compound formulas, as well as its substitutes, quality control, formulation study, compound prescription, pharmacological mechanism and clinical research. The aim of this article is to provide a valuable reference for future developments and studies of Calculus Bovis.

Dynamic observation of asthma model induced by allergen in mice from acute inflammation to chronic remodeling / 中国病理生理杂志

AIM:To dynamically observe and compare the relative changes of the indexes from the process of acute inflammation to chronic remodeling in asthmatic mice induced by ovalbumin ( OVA) .METHODS:Female BALB/c mice (n=60) were randomly divided into normal control group and asthma group .The mice in asthma group were sensi-tized and challenged by OVA , while the mice in normal group received equal volume of normal saline ( NS) .The challenge was performed for 3 consecutive days from the 21th day to observe the response of acute inflammation , and then the mice in different groups were challenged once per week for 5 weeks.Detailed comparisons of the dynamic changes of cell infiltra-tion, cytokine expression and airway remodeling were conducted .RESULTS:Compared with NS group , the mice in OVA group showed a predominantly eosinophilic infiltration into the airway lumen , increased production of Th 2-type cytokines , secretion of epithelial mucus and deposition of subepithelial collagen .In OVA challenge groups , the levels of inflammatory cells and inflammatory factors were remarkably higher in 24 d group, whereas the most obvious changes of goblet cell hyper-plasia and airway remodeling were observed in 52 d group.CONCLUSION:Acute asthma model is sufficiently induced by 3 consecutive days of OVA challenge protocol , which is accompanied with high levels of inflammatory cells and inflammato-ry factors.The OVA challenge protocol once per week for 5 weeks could induce a chronic asthma model with obvious airway remodeling .