Bioethnopharmaceutical analytical pharmacology (BAP) means to study Chinese herbal compound formula (CHCF) from the aspacts of in vivo and in vitro efficacy,pharmacodynamics,quality control and plant chemistry,guided by the CHCF absorbed bioactive compounds (ABC) analyses.The form of BAP is performed by comparing ABC efficacy with mother formula efficacy.Meanwhile,it must follow the principle which the ABC dose should be equal to the mother formula content or the blood drug concentration.In this study,the hypothesis was put forward to clarify the thoughts,assumptions and expected results,which uncovered the multiple cardioprotective mechanism of Dangui-Buxue Tang (DBT) for ischemic heart disease.BAP is expected to guide the development of further experiments for providing a better thought for the research over CHCF.

Objectives To study the psychological characteristics of children with attention deifcit hyperactivity disorder (ADHD). Methods Human ifgure drawings (HFD) as a projective technique was performed in 107 children with ADHD aged (8.69 ±1.48) years old (6.5~11.5 years), and 107 matched healthy children. Characteristic features of the HFDs in children with ADHD, and correlations between emotional indicators of the HFDs and factors of child behavior checklist (CBCL) were assessed and com-pared with the control group. Results Signiifcant differences in emotional index were found between ADHD and healthy chil-dren, including bad body composition, short arms, long arms, big ifgure, omission of hands and omission of feet (P<0.05). Nega-tive correlation was found between the HFDs emotional index and the factors of activity and social ability in the CBCL (P<0.05), but a positive correlation was found between the HFDs emotional index and behavioral problem (P<0.05). Conclusion Human ifgure drawings can relfect some emotional characteristics of children with ADHD, and when combined with CBCL, it can pro-vide evidence for diagnosis and intervention for ADHD.

Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.

Objective To evaluate clinicopathologic features and prognosis of primary gastric anaplastic large cell lymphomas (ALCL).Methods Clinical data and parafiin blocks of 4 patients diagnosed with primary gastric ALCL were obtained. The diagnosis of all cases was based on the criteria of WHO classification of hematolymphoid neoplasm.Furthermore,chromosomal rearrangement involving ALK gene was detected by interphase fluorescence in situ hybridization (FISH) and Epstein-Barr virus (EBV) status was determined by in situ hybridization(ISH) for EBV-encoded small RNAs (EBERs).Results The patients (3 males and 1 female) were from 27 to 87 years old, with a median age of 58.5 years. All the four cases presented with a solitary ulcerative mass in stomach. Morphologically, the normal architecture of gastric wall was effaced by the diffuse infiltration of tumor cells in which the characteristic hallmark cells were easily identified.The tumor cells of all cases showed a consistently strong expression of LCA and CD30,and CD3e was expressed in 3 of the 4 cases.Both ALK expression and ALK gene rearrangements were negative in all cases.Two cases underwent total or partial gastrectomy followed by CHOP chemotherapy. Another one patient was treated with chemotherapy and autologous stem cell transplantation. None of these 3 patients developed a relapse or progression till the last follow-up on Nov 30,2011. While the rest one patient refused to take any treatment and died 20 months after diagnosis. Conclusions Primary gastric ALCL is very rare and usually ALK negative. Its pathologic features as well as the clinical outcome are quite similar to the ALK negative ALCL from other sites, except the more frequently positive CD3e Early diagnosis and proper therapy are of great significance to the prognosis.

Objective To study the clinicopathologic features and explore the potential prognostic factors of extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma).Methods 86 patients diagnosed with MALT lymphoma were retrieved and divided into 2 subgroups according to the location of tumor,namely gastric (32 cases) and non-gastric (54 cases) subgroup.Histological characteristics were reassessed on hematoxylin-eosin staining slides,and immunophenotypic features were determined by immunohistochemical staining.Interphase fluorescence in-situ hybridization (FISH) was carried out to detect the cytogenetic abnormalities.Results There were no significant difference of clinical behavior,histology,and immunophenotype between gastric and non-gastric subgroups.In addition,FISH detected t(14;18) in 9 ％ (4/45) and t(11;18) in 12 ％ cases (6/50) respectively.Among the 20 cases treated with H pylori (HP) eradication,a significantly lower remission rate was observed in cases with bcl-10 (20 ％) nuclear expression or those harboring t(11;18) (33 ％) compared with those in the negative group (73 ％,91％)(x2 =3.842 and 4.639,P =0.035 and 0.031,respectively).For the 35 non-gastric patients with follow-up data,males (35 ％) and patients older than 60 yrs (25 ％) tended to have a lower remission rate as compared to females (60 ％) and those younger than 60 yrs (63 ％) (x2 =3.905 and 7.373,P =0.048 and 0.007,respectively).Moreover,progression-free survival rate was significantly lower in patients with higher stage (ⅢⅣ) (25 ％) and without t(14;18) (50 ％) than that in patients with stage Ⅰ-Ⅱ (52 ％) and with t(14;18)(75 ％).The differences had statistical significance (x2 =4.207 and 4.363,P =0.040 and 0.037,respectively).Conclusion MALT lymphoma in general is an indolent B-cell non-Hodgkin’ s lymphoma which more frequently occurs in the elderly people.Differences in the response to treatment or the prognosis are observed between gastric and non-gastric MALT lymphoma patients.

Objective To observe the influence of Qizhi Yifei containing serum on the expression of MMP-9 and TIMP-1 mRNA in lung fibroblasts, and explore its mechanism. Methods Trypsin digestion method was used to extract fibroblasts from lung tissue in rats. All fibroblasts were cultured and trained to the fourth generation. Then they were randomly divided into control group, model group and drug serum group. The model group and drug serum group were firstly treated by DMEM with 0.002 5 μg/mL TGF-β1. The control group was treated by DMEM only. The control group and model group were then treated by DMEM with blank drug serum in concentration of 5%, and the drug serum group was treated by DEME with Qiahi Yifei containing serum in same concentration. After 48 and 72 hours, RT-PCR was performed to test the expression of MMP-9 mRNA and TIMP-1 mRNA of each group. Results After 48 hours, MMP-9 and TIMP-1 mRNA were significantly increased in model group and drug serum group compared with control group. There was no difference between model group and drug serum group. After 72 hours, MMP-9 mRNA was up-regulated in model group and was decreased in drug serum group compared with control group. There was no significant difference among the three groups on the expression of TIMP-1 mRNA. Conclusion Qizhi Yifei containing serum can decrease the up-regulated expression of MMP-9 mRNA in lung fibroblasts stimulated by TGF-β1.

Objective To monitor the autophagy in osteosarcoma cells by constructing three rLC3B fusion expression vectors,respectively.Methods Rat LC3B gene sequence was amplified by PCR and cloned into pEGFP-C 1 and pmCherry-C1 to construct the fusion expression vector of pEGFP-rLC3B and pmCherry-rLC3B.Subsequently,the EGFP-rLC3B sequence was obtained by PCR with the pEGFP-rLC3B as a template,and cloned into pmCherry-C 1,so the pmCherry-EGFP-rLC3B fusion expression vector was constructed.Three plasmids were transfected into U-2OS cells,and the starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A1 was used to inhibit autophagy,to verify the above plasmids' function in autophagy detection by laser scanning confocal microscopy.Western blot was used to detect the endogenous LC3B and exogenous EGFPrLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B,and to verify the correct expression of exogenous rLC3B and their function of autophagy detection.Finally,cleaved free EGFP was detected by western blot to evaluate the level of autophagic degradation.Results Three fusion expression vectors were constructed successfully through sequencing and restriction enzyme digestion validation.The starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A 1 was used to inhibit autophagy in transfected U-2OS cells.Clear autophagosomes and autolysosomes were observed by laser scanning confocal microscopy.Endogenous LC3B and exogenous EGFP-rLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B were detected through western blot.Finally,western blot verified that the expression of cleaved free EGFP was significantly up-regulated with the increase of starvation time.12 h group increased 1.05 times than the control group and 24 h group increased 1.56 times,showing that the levels of autophagic degradation increased.Conclusion EGFP-rLC3B can be used to detect autophagosome and evaluate the level of autophagic degradation.mCherry-rLC3B can be used to detect autophagosome and autolysosome,but can't distinguish autophagosome from autolysosome.The pmCherry-EGFP-rLC3B has an advantage in the detection of autophagic flux which can distinguish autophagosome from autolysosome.

OBJECTIVETo study chemical constituents contained in Desmodium caudatum.

METHODThe chemical compounds were separated by using such chromatographic methods as macroporous resin, Sephadex LH-20, ODS and normal phase silicagel column, and their structures were identified by spectroscopic data analysis.

RESULTFifteen compounds were separated and identified as stigmasterol (1), beta-sitosterol (2), citrusinol (3), hibiscone A (4), yukovanol (5), kenusanone I (6), neophellamuretin (7), desmodol (8), erythrotriol (9), hibiscone D (10), kaempferol (11), 8-prenylquercetin (12), leachianone G (13), 5,7,4'-trihydroxy-dihydroflavonol (14), and 4H-1-benzopyran-4-one, 2-(3,4-dihydroxyphenyl) -2, 3-dihydro-3,5,7-trihydroxy-8-( 3-methyl-2-butenyl) -, (2R-trans)-(9CI) (15).

CONCLUSIONAll of the compounds were separated from D. caudatum for the first time except compound 8.

Objective: To observe the effects of different concentrations and doses of urea on the proliferation and apoptosis of human hemangioma endothelial cells, in order to provide evidence for the further mechanism study of urea in the treatment of hemangioma. Methods: Human hemangioma endothelial cells (HemECs) and normal endothelial cells (VE) were cultured in vitro. Cell viability was detected by CCK-8 after invention with different concentrations(40%, 50%, 60%, 70%) and doses(3, 6, 9 μl/ml) of urea. The apoptosis of HemECs was detected by flow cytometry dual-dye and propidium lodide single dye. Results: The viability of HemECs was significantly lower than that of VE under different concentrations and doses of urea (P<0.05). The inhibition rate of 40% urea on HemECs increased with the increase of urea dose (P<0.05), and the inhibition effect was most obvious at 4 h and 12 h. The apoptosis of HemECs increased in a time and dosage dependent manner with the treatment of 40% urea. High dose(9 μl/ml)of 40% urea significantly promoted the apoptosis of HemECs(P<0.05). Conclusions Low dose of 40% urea significantly inhibited the proliferation of HemECs, and had no significant effect on VE. However, high doses of urea promoted apoptosis of HemECs.

Objective: To establish a set of rules for autoverification of blood analysis, in order to provide a way to validate autoverification rules for different analytical systems, which can ensure the accuracy of test results as well as shorten turnaround time (TAT) of test reports. Methods: A total of 34 629 EDTA-K2 anticoagulated blood samples were collected from multicenter cooperative units including the First Hospital of Jinlin University during January 2017 to November 2017. These samples included: 3 478 cases in Autoverification Establishment Group, including 288 cases for Delta check rules; 5 362 cases in Autoverification Validation Group, including 2 494 cases for Delta check; 25 789 cases in Clinical Application Trial Group. All these samples were analyzed for blood routine tests using Sysmex XN series automatic blood analyzers.Blood smears, staining and microscopic examination were done for each sample; then the clinical information, instrument parameters, test results and microscopic results were summarized; screening and determination of autoverification conditions including parameters and cutoff values were done using statistical analysis. The autoverification rules were input into Sysmex Laboman software and undergone stage Ⅰ validation using simulated data, and stage Ⅱ validation for post-analytical samples successively. True negative, false negative, true positive, false positive, autoverification pass rate and passing accuracy were calculated. Autoverification rules were applied to autoverification blood routine results and missed detection rates were validated, and also data of autoverification pass rate and TAT were obtained. Results: (1)The selected autoverification conditions and cutoff values included 43 rules involving WBC, RBC, PLT, Delta check and abnormal characteristics. (2)Validation of 3 190 cases in Autoverification Establishment Group showed the false negative rate was 1.94%(62/3 190)(P<0.001), autoverification pass rate was 76.74%, passing accuracy was 97.47%; Validation of 2 868 cases in Autoverification Validation Group, the false negative rate was 3.38%(97/2 868)(P=0.002), autoverification pass rate was 42.26%, passing accuracy was 92.00%; Validation of Delta check on 288 cases in Autoverification Establishment Group and 2 494 cases in Autoverification Validation Group showed the false negative rates were respectively 1.39% and 2.61%(P<0.001). (3)Three hospitals adopted these rules of autoverification for 25 789 blood routine samples, and found that the average TAT of blood routine test reports were shortened by 24min, 32min and 7min respectively, the rate of samples reported within 30min were elevated by 33%, 53% and 7%. The autoverification pass rates were 72%-74%. Conclusions The application of this set of 43 autoverification rules in blood sample analysis can ensure test quality while shortenTAT and improve work efficiency. It is worth pointing out that for the same analytical systems in this research, validation is necessary before application of this set of rules, and periodic validation is required during application to make necessary adjustment; for different analytical systems, as this research provide a way to establish autoverification rules for blood routine tests.Clinical labs may establish their own suitable autoverification rules on the basis of technological parameters. (Chin J Lab Med, 2018, 41: 601-607)