Current status of diagnosis and treatment of chronic lymphocytic leukemia in China: A national multicenter survey research / 中华血液学杂志

Objective: To understand the current status of diagnosis and treatment of chronic lymphocytic leukemia (CLL) /small lymphocytic lymphoma (SLL) among hematologists, oncologists, and lymphoma physicians from hospitals of different levels in China. Methods: This multicenter questionnaire survey was conducted from March 2021 to July 2021 and included 1,000 eligible physicians. A combination of face-to-face interviews and online questionnaire surveys was used. A standardized questionnaire regarding the composition of patients treated for CLL/SLL, disease diagnosis and prognosis evaluation, concomitant diseases, organ function evaluation, treatment selection, and Bruton tyrosine kinase (BTK) inhibitor was used. Results: ①The interviewed physicians stated that the proportion of male patients treated for CLL/SLL is higher than that of females, and the age is mainly concentrated in 61-70 years old. ②Most of the interviewed physicians conducted tests, such as bone marrow biopsies and immunohistochemistry, for patient diagnosis, in addition to the blood test. ③Only 13.7% of the interviewed physicians fully grasped the initial treatment indications recommended by the existing guidelines. ④In terms of cognition of high-risk prognostic factors, physicians' knowledge of unmutated immunoglobulin heavy-chain variable and 11q- is far inferior to that of TP53 mutation and complex karyotype, which are two high-risk prognostic factors, and only 17.1% of the interviewed physicians fully mastered CLL International Prognostic Index scoring system. ⑤Among the first-line treatment strategy, BTK inhibitors are used for different types of patients, and physicians have formed a certain understanding that BTK inhibitors should be preferentially used in patients with high-risk factors and elderly patients, but the actual use of BTK inhibitors in different types of patients is not high (31.6%-46.0%). ⑥BTK inhibitors at a reduced dose in actual clinical treatment were used by 69.0% of the physicians, and 66.8% of the physicians had interrupted the BTK inhibitor for >12 days in actual clinical treatment. The use of BTK inhibitors is reduced or interrupted mainly because of adverse reactions, such as atrial fibrillation, severe bone marrow suppression, hemorrhage, and pulmonary infection, as well as patients' payment capacity and effective disease progression control. ⑦Some differences were found in the perceptions and behaviors of hematologists and oncologists regarding the prognostic assessment of CLL/SLL, the choice of treatment options, the clinical use of BTK inhibitors, etc. Conclusion: At present, a gap remains between the diagnosis and treatment of CLL/SLL among Chinese physicians compared with the recommendations in the guidelines regarding the diagnostic criteria, treatment indications, prognosis assessment, accompanying disease assessment, treatment strategy selection, and rational BTK inhibitor use, especially the proportion of dose reduction or BTK inhibitor discontinuation due to high adverse events.

Progress on clinical trials of common gastrointestinal cancer drugs in China from 2012 to 2021 / 中华肿瘤杂志

Objective: Systematically summarize the research progress of clinical trials of gastric cancer oncology drugs and the overview of marketed drugs in China from 2012 to 2021, providing data and decision-making evidence for relevant departments. Methods: Based on the registration database of the drug clinical trial registration and information disclosure platform of Food and Drug Administration of China and the data query system of domestic and imported drugs, the information on gastric cancer drug clinical trials, investigational drugs and marketed drugs from January 1, 2012 to December 31, 2021 was analyzed, and the differences between Chinese and foreign enterprises in terms of trial scope, trial phase, treatment lines and drug type, effect and mechanism studies were compared. Results: A total of 114 drug clinical trials related to gastric tumor were registered in China from 2012 to 2021, accounting for 3.7% (114/3 041) of all anticancer drug clinical trials in the same period, the registration number showed a significant growth rate after 2016 and reached its peak with 32 trials in 2020. Among them, 85 (74.6%, 85/114) trials were initiated by Chinese pharmaceutical enterprise. Compared with foreign pharmaceutical enterprise, Chinese pharmaceutical enterprise had higher rates of phase I trials (35.3% vs 6.9%, P=0.001), but the rate of international multicenter trials (11.9% vs 67.9%, P<0.001) was relatively low. There were 76 different drugs involved in relevant clinical trials, of which 65 (85.5%) were targeted drugs. For targeted drugs, HER2 is the most common one (14 types), followed by PD-1 and multi-target VEGER. In the past ten years, 3 of 4 marketed drugs for gastric cancer treatment were domestic and included in the national medical insurance directory. Conclusions: From 2012 to 2021, China has made some progress in drug research and development for gastric carcinoma. However, compared with the serious disease burden, it is still insufficient. Targeted strengthening of research and development of investment in many aspects of gastric cancer drugs, such as new target discovery, matured target excavating, combination drug development and early line therapy promotion, is the key work in the future, especially for domestic companies.

Analysis of Clinical Characteristics of JAK2 V617F and BCR-ABL Double-Mutant Myeloproliferative Neoplasms / 中国实验血液学杂志

OBJECTIVE@#To analyze the disease types, clinical manifestations, efficacy and outcome of JAK2 V617F and BCR-ABL double-mutant myeloproliferative neoplasms (MPN), and provide a reference for the diagnosis, treatment and prognosis of MPN.@*METHODS@#The clinical characteristics, diagnosis, therapeutic efficacy and outcome of JAK2 V617F and BCR-ABL double-mutant MPN were analyzed comprehensitively by combining a clinical case diagnosed and treated in our hospital with literature cases from CNKI and PubMed databases.@*RESULTS@#A total of 38 related literatures were retrieved from the two databases by searching "JAK2 V617F" and "BCR-ABL" as key words from 1990 to 2019, and 59 cases were involved. Among all the 60 cases, 41 were males (68.3%) with a median age of 61 (32-77) years old, while 19 were females (31.7%) with a median age of 58 (21-82) years old. The BCR-ABL fusion gene and JAK2 V617F mutation were found simultaneously in 21 cases (35%), 19 cases (31.7%) with JAK2 V617F mutation were found during the treatment of Philadelphia chromosome (Ph)-positive chronic myelogenous leukemia (CML). Ph@*CONCLUSION@#As cases of BCR-ABL and JAK2 V617F double-mutant MPN are reported, simultaneous detection of JAK2 V617F mutation and BCR-ABL fusion gene in MPN patients is necessary to avoid misdiagnosis and missed diagnosis.

Hand Function Rehabilitation Management Program after Burn: Based on Knowledge to Action Framework / 中国康复理论与实践

Objective:To construct a management program of hand function rehabilitation for adult inpatients after hand burn. Methods:Literatures of clinical practice guideline, evidence summary, systematic review and review were searched from OVID JBI, Cochranel Library, EBSCO, PubMed、EMbase、FEBM, CNKI, Wanfang Data and CBM, and so on. They were screened, evaluated, extracted evidence, found hinder factors following the steps of Knowledge to Action (KTA). Finally, a hand function rehabilitation management program was constructed. Results:Ten literatures were included. The management program included hand function rehabilitation intervention program and implementation procedures. Conclusion:Based on KTA framework, the hand function rehabilitation management program after burn can be used in clinic.

Hand Function Rehabilitation Management Program after Burn: Based on Knowledge to Action Framework / 中国康复理论与实践

Objective:To construct a management program of hand function rehabilitation for adult inpatients after hand burn. Methods:Literatures of clinical practice guideline, evidence summary, systematic review and review were searched from OVID JBI, Cochranel Library, EBSCO, PubMed、EMbase、FEBM, CNKI, Wanfang Data and CBM, and so on. They were screened, evaluated, extracted evidence, found hinder factors following the steps of Knowledge to Action (KTA). Finally, a hand function rehabilitation management program was constructed. Results:Ten literatures were included. The management program included hand function rehabilitation intervention program and implementation procedures. Conclusion:Based on KTA framework, the hand function rehabilitation management program after burn can be used in clinic.

Puerarin attenuates angiotensin II-induced cardiac fibroblast proliferation via the promotion of catalase activity and the inhibition of hydrogen peroxide-dependent Rac-1 activation / 中国天然药物

The aims of the present study were to evaluate the effects of puerarin on angiotensin II-induced cardiac fibroblast proliferation and to explore the molecular mechanisms of action. Considering the role of HO in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation, we hypothesized that modulating catalase activity would be a potential target in regulating the redox-sensitive pathways. Our results showed that the activation of Rac1 was dependent on the levels of intracellular HO. Puerarin blocked the phosphorylation of extracellular regulated protein kinases (ERK)1/2, abolished activator protein (AP)-1 binding activity, and eventually attenuated cardiac fibroblast proliferation through the inhibition of HO-dependent Rac1 activation. Further studies revealed that angiotensin II treatment resulted in decreased catalase protein expression and enzyme activity, which was disrupted by puerarin via the upregulation of catalase protein expression at the transcriptional level and the prolonged protein degradation. These findings indicated that the anti-proliferation mechanism of puerarin was mainly through blocking angiontensin II-triggered downregulation of catalase expression and HO-dependent Rac1 activation.

Puerarin attenuates angiotensin II-induced cardiac fibroblast proliferation via the promotion of catalase activity and the inhibition of hydrogen peroxide-dependent Rac-1 activation / 中国天然药物

The aims of the present study were to evaluate the effects of puerarin on angiotensin II-induced cardiac fibroblast proliferation and to explore the molecular mechanisms of action. Considering the role of HO in nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activation, we hypothesized that modulating catalase activity would be a potential target in regulating the redox-sensitive pathways. Our results showed that the activation of Rac1 was dependent on the levels of intracellular HO. Puerarin blocked the phosphorylation of extracellular regulated protein kinases (ERK)1/2, abolished activator protein (AP)-1 binding activity, and eventually attenuated cardiac fibroblast proliferation through the inhibition of HO-dependent Rac1 activation. Further studies revealed that angiotensin II treatment resulted in decreased catalase protein expression and enzyme activity, which was disrupted by puerarin via the upregulation of catalase protein expression at the transcriptional level and the prolonged protein degradation. These findings indicated that the anti-proliferation mechanism of puerarin was mainly through blocking angiontensin II-triggered downregulation of catalase expression and HO-dependent Rac1 activation.

Construction of Ontological Modeling for Core Concepts of TCM Sub-domain / 中国中医药信息杂志

Objective In view of the inconsistency between the concepts of TCM in the field of TCM information intersection, the confusion of domain knowledge expression caused by conceptual cross and term ambiguity, and taking TCM sub-domain as a sample, knowledge representation and ontological modeling of core concepts are realized. Methods Through collecting and combing 2015 version of Pharmacopoeia of People's Republic of China (the first volume), Coding Rules for Chinese Medicines and Their Codes, Chinese Materia Medica, TCM Dictionary, Science of Chinese Materia Medica, Clinical Science of Chinese Materia Medica and other TCM knowledge authority source data, based on ontological conceptual modeling method, this article confirmed the definition and connotation of Chinese materia medica, determined the concept properties, clarified the relationship between concepts, and initially completed the ontological modeling for core concepts of TCM sub-domain. Results The ontological modeling of key concepts such as "TCM", "Chinese materia medica", "TCM decoction pieces" and "TCM patent medicines" was completed. Conclusion The preliminary construction of TCM related core conceptual model is realized, which can provide reference for expressions and sharing of TCM knowledge in other sub-domains.

Correlation of Gemin 3 SNP in the microRNA Biosynthesis Pathway with Risk and Prognosis of Patients with Non-Hodgkin's Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the correlation of single nucleotide polymorphisms (SNP) of Gemin 3(rs197412) in the miRNA biosynthesis with NHL cancer risk and overall prognosis.</p><p><b>METHODS</b>miR-SNP were genotyped using PCR-ligase detection reaction(LAR, LCR) in NHL group of 230 non-Hodgkin lymphoma (NHL) patients and in control group of 120 healthy persons. The survival curves were drawn using the Kaplan-Meier method, and comparisons between the curves were made using the log-rank test. Multivariate survival analysis was performed by using a Cox proportional hazards model.</p><p><b>RESULTS</b>The rs197412 genotype distribution difference was not statistically significant, in NHL and control group; the survival time of patients carrying the rs197412 TT genotype was significantly longer than that of the patients carrying the CC+CT genotype (P=0.007). In addition, rs197412 was independent from the survival of NHL patients by multivariate analysis (RR: 2.138,95% CI: 1.303-3.508, P<0.01).</p><p><b>CONCLUSION</b>The single nucleotide polymorphisms of Gemin 3 (rs197412) in the miRNA processing are not related with NHL risk, but that may affect NHL survival.</p>

Selection of syndrome concepts and establishment of relationship between syndrome concepts in Traditional Chinese Medicine Clinical Terminological System version2.0 / 中华医学图书情报杂志

After the methods for selection of syndrome concepts and establishment of relationship between syn-drome concepts in Traditional Chinese Medicine Clinical Terminological System version2.0 were described, the principles and specific methods for the selection of syndrome concepts and establishment of relationship between syn-drome concepts in Traditional Chinese Medicine Clinical Terminological system version2.0 were studied according to its general revision principles in the light of relevant standards in traditional Chinese medicine, various editions of teaching materials and medical records, the advantages of descriptive definition and logical definition of syndrome concepts,and the establishment of relationship between syndrome concepts derived from syndrome concepts were elaborated.

Advances of studies on pathogenesis and management of paroxysmal nocturnal hemoglobinuria / 中国实验血液学杂志

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal disorder of hematopoiesis due to the inactivation of PIG-A gene. However, the presence of mutant PIG-A gene in a group of hematopoietic cells is not enough for the development of PNH, immunologic injury and apoptotic effects are considered to play an important role in clonal expansion. Knowledge of the molecular mechanisms leading to PNH has substantially increased in the past decades, which remarkably advances the diagnostic modalities and treatment approaches of patients with PNH. Though great progress has been made because of targeted therapy method, the challenges are still ahead. In this review the advances of studies on mechanism, laboratorial diagnosis and therapeutic protocols of PNH are summarized.

Pharmacokinetics of enteric-coated mycophenolate sodium in Chinese renal transplantation recipients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Mycophenolic acid (MPA) as an anti-proliferative immune-suppressive agent is used in the majority of immunosuppressive regimens in solid organ transplantation. This study aimed to investigate the pharmacokinetic (PK) characteristics of enteric-coated mycophenolate sodium (EC-MPS) and area under the curve (AUC) from 0 to 12 hours with limited sampling strategies (LSSs) in Chinese renal transplant recipients.</p><p><b>METHODS</b>This study was conducted in 10 Chinese renal transplant patients receiving living donor and treated with EC-MPS, cyclosporine, and corticosteroids. MPA concentrations were measured by enzyme multiplied immunoassay technique (EMIT). Whole 12-hour PK profiles were obtained on Day 4 after operation. LSSs with jackknife technique, multiple stepwise regression analysis, and Bland-Altman analysis were developed to estimate MPA AUC.</p><p><b>RESULTS</b>The mean maximum plasma concentration, the mean time for it to reach peak (T(max)), and the mean MPA AUC were (11.38 ± 2.49) mg/L, (4.85 ± 3.32) hours, and (63.19 ± 13.54) mg×h×L(-1), respectively. Among the 10 profiles, MPA AUC of four patients was significantly higher than that of the other six patients, and the corresponding T(max) was significantly longer than that of the other six patients. No patient exhibited a second peak caused by enterohepatic recirculation. The best models were as follows: 27.46 + 0.94C(3) + 3.24C(8) + 2.81C(10) (r(2) = 0.972), which was used to predict AUC of fast metabolizer with a mean prediction error (MPE) of -0.21% and a mean absolute prediction error (MAE) of 2.59%; 36.65 + 3.08C(8) + 5.30C(10) - 4.04C(12) (r(2) = 0.992), which was used to predict AUC of slow metabolizer with a MPE of 0.58% and a MAE of 1.95%.</p><p><b>CONCLUSIONS</b>The PKs of EC-MPS had a high variability among Chinese renal transplant recipients. The preliminary PK data indicated the existence of slow and fast metabolizer. These findings may be associated with the enterohepatic recirculation.</p>

Detoxication effect of water-soluble imprinted cross-linked chitosan on depleted uranium induced toxicity to renal cells / 药学学报

To investigate whether a series of water-soluble cross-linked chitosan derivates synthesized in the guide of imprinting technology could be used as a uranium chelating agent to protect cells exposed to depleted uranium (DU), the imprinted chitosan derivates with high UO2(2+) chelating ability were screened, and cell model of human renal proximal tubule epithelium cells (HK-2) exposed to DU (500 micromol.L-1) was built, chitosan derivates (400 mg.L-1 ) was added to test group and diethylenetriaminepentaacetic acid (DTPA, 50 mg.L-1) was added to positive control group. The results showed that three Cu2+ imprinted chitosan derivates had higher uranium chelating ability (>49 microg.mg-1) than chitosan and non-imprinted chitosan derivates. Compared to the cells exposed to DU only, survival of cells in group added chitosan derivates rose up significantly (increased from 57.3% to 88.7%, and DTPA to 72.6%), and DU intracellular accumulation decreased, membrane damage and DNA damage also eased. Among the imprinted chitosan derivates, Cu2+ imprinted penta dialdehyde cross-linked carboxymethyl chitosan (Cu-P-CMC) was the best, and better than DTPA. From ultrastructure observation, the DU precipitates of test group added Cu-P-CMC were most grouped in a big hairy clusters in a string together outside cells. It is possible that the DU-chitosan derivates precipitates are too big to enter into cells, and from this way, the DU uptake by cells decreased so as to detoxication.

Prediction and bioinformatics analysis of human gene expression profiling regulated by amifostine / 中国实验血液学杂志

Objective of this study was to perform bioinformatics analysis of the characteristics of gene expression profiling regulated by amifostine and predict its novel potential biological function to provide a direction for further exploring pharmacological actions of amifostine and study methods. Amifostine was used as a key word to search internet-based free gene expression database including GEO, affymetrix gene chip database, GenBank, SAGE, GeneCard, InterPro, ProtoNet, UniProt and BLOCKS and the sifted amifostine-regulated gene expression profiling data was subjected to validity testing, gene expression difference analysis and functional clustering and gene annotation. The results showed that only one data of gene expression profiling regulated by amifostine was sifted from GEO database (accession: GSE3212). Through validity testing and gene expression difference analysis, significant difference (p < 0.01) was only found in 2.14% of the whole genome (460/192000). Gene annotation analysis showed that 139 out of 460 genes were known genes, in which 77 genes were up-regulated and 62 genes were down-regulated. 13 out of 139 genes were newly expressed following amifostine treatment of K562 cells, however expression of 5 genes was completely inhibited. Functional clustering displayed that 139 genes were divided into 11 categories and their biological function was involved in hematopoietic and immunologic regulation, apoptosis and cell cycle. It is concluded that bioinformatics method can be applied to analysis of gene expression profiling regulated by amifostine. Amifostine has a regulatory effect on human gene expression profiling and this action is mainly presented in biological processes including hematopoiesis, immunologic regulation, apoptosis and cell cycle and so on. The effect of amifostine on human gene expression need to be further testified in experimental condition.

Relationship between proximal-tip location and partial necrosis in distally based sural neuro fasciocutaneous flap: an analysis of 157 flaps / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the influence of proximal-tip location on partial necrosis in distally based sural neuro fasciocutaneous flap.</p><p><b>METHODS</b>From April 2001 to May 2009,157 distally based sural neuro fasciocutaneous flaps were conducted to repair the soft tissue defect in distal region of lower leg, ankle and feet in 153 patients. Date of the flaps and the patients were retrospectively analyzed. From the tip of lateral malleolus to the popliteal crease, posterior aspect of the lower leg was equally divided into 9 regions that were 1st to 9th region from inferiorly to superiorly, respectively. The flaps were divided into 2 groups: survival group (including uneventfully survived flaps, flaps with distally epidermal necrosis and with wound dehiscence) and partial necrosis group. Based on the location of the proximal tip of flaps, the flaps were stratified into 4 groups: flaps with the proximal tip locating in the 6th or lower region (group A), the 7th region (group B), the 8th region (group C) and the 9th region (group D). Harvesting the flaps started from exploring the perforator of peroneal vessel in the adipofascial pedicle, then the flaps were elevated retrogradely.</p><p><b>RESULTS</b>Of the 157 flaps, 125 survived uneventfully,8 showed distal epidermal necrosis,wound dehiscence occurred in 6 flaps, 18 flaps (11.5%) showed distal partial necrosis. Partial necrosis occurred in zero of 19 flaps in group A (0), 1 of 44 flaps in group B (2.3% ), 7 of 62 flaps in group C (11.3% ) and 10 of 32 flaps in group D (31.3% ). The differences in partial necrosis rate between group A and group B , group B and group C, were not statistically significant (P > 0.05). Partial necrosis rate was higher in group D than in group C (P = 0.012), it was lower in group A + group B (1.6%) than in group C + group D (18. 1% ) (P = 0. 001).</p><p><b>CONCLUSIONS</b>Distally based sural neuro fasciocutaneous flap can survive reliably when the proximal tip of flap is not beyond the junction between lower 7/9 and upper 2/9 of the lower leg, whereas probability of partial necrosis occurring in the flap increase significantly when the proximal tip of flap locates in upper 1/9 of the lower leg.</p>

Application of Fourier amplitude sensitivity test in Chinese healthy volunteer population pharmacokinetic model of tacrolimus / 药学学报

In this study, we evaluated the influence of different variance from each of the parameters on the output of tacrolimus population pharmacokinetic (PopPK) model in Chinese healthy volunteers, using Fourier amplitude sensitivity test (FAST). Besides, we estimated the index of sensitivity within whole course of blood sampling, designed different sampling times, and evaluated the quality of parameters' and the efficiency of prediction. It was observed that besides CL1/F, the index of sensitivity for all of the other four parameters (V1/F, V2/F, CL2/F and k(a)) in tacrolimus PopPK model showed relatively high level and changed fast with the time passing. With the increase of the variance of k(a), its indices of sensitivity increased obviously, associated with significant decrease in sensitivity index for the other parameters, and obvious change in peak time as well. According to the simulation of NONMEM and the comparison among different fitting results, we found that the sampling time points designed according to FAST surpassed the other time points. It suggests that FAST can access the sensitivities of model parameters effectively, and assist the design of clinical sampling times and the construction of PopPK model.

Gene expression profile analysis of T lymphocytes involved in pathogenesis of severe aplastic anemia by using bioinformatics method as a novel way of drug screening / 中国实验血液学杂志

This study was aimed to explore the gene expression profile characteristics of T lymphocytes involved in pathogenesis of severe aplastic anemia (SAA) and to predict putative curative drugs for SAA by using biological principle of similarity contrast of gene expression profiles between drugs and diseases. SAA and T lymphocyte were used as key words to search gene expression datasets related to pathogenesis of SAA in public Gene Expression Omnibus (GEO) of NCBI. After significance test, gene expression profiling involved in pathogenesis of SAA were screened and applied to cluster analysis. And then SAA-related gene expression profiles were thrown into pharmacological gene expression datasets of 3000 candidate drugs for similarity analysis and significantly negative correlation was used as a screening criterion for selecting putative curative drugs of SAA. The results showed that only one gene expression dataset was found out, i.e. GSE3807. Computational bioanalysis identified a total of 515 candidate genes of T lymphocyte involved in pathogenesis of SAA, whose expression level exceeded more than 2-fold. Among them, 202 genes were upregulated and 313 genes were downregulated. Cluster analysis showed that those genes belonged to different pathways, including nucleic acid metabolic process, ubiquitin-dependent protein catabolic process, Golgi apparatus protein transport, protein phosphorylation and immunoglobin/major histocompatibility complex. Similarity analysis of gene expression profiles of SAA and drugs showed that hydroxycamptothecin and metformin might have a potential therapeutic efficacy on SAA. It is concluded that by means of novel bioinformatics method, gene expression profiling combined with similarity analysis between disease-related gene expression and pharmacological gene expression profiles may be a novel way of drug screening for SAA.

Cloning of ID4 gene expression regulation promoter and subcloning of recombinant ID4 promoter luciferase reporter / 中国实验血液学杂志

<p><b>UNLABELLED</b>The present study was aimed to clone ID4 gene promoter and upstream regulatory region, and to construct a series of recombinant promoter-luciferase reporter for exploring the mechanism of ID4 gene expression regulation.</p><p><b>METHODS AND RESULTS</b>the upstream 5' flanking sequence of 2242 bp from transcriptional start site (TSS) and downstream 5' non-coding region of 212 bp on ID4 gene were searched out and downloaded from human genome databank of NCBI using whole length of ID4 gene cDNA as a probe; On-line promoter analysis softwares, including TESS and Genomax, were employed to analyze the characteristics of ID4 gene promoter and upstream regulatory elements. Then, based on the analytic results, PCR primers were designed and synthesized. Segmental amplification method was adopted to obtain two fragments of 1829 bp and 784 bp. The two fragments were inserted into the plasmid pGEM-T, transformed into TOP10 competent E. coli., and positive recombinants were screened respectively. Subsequently, restriction enzymes KpnI/NheI and KpnI/EcoRI were used to digest the above-mentioned two plasmids pGEM-T and pGL3, and ligation was completed by T4 DNA ligase. After transformation to TOP10 competent E. coli. and screening of positive colonies, the basic recombinant ID4 gene promoter-pGL3 was successfully constructed. KpnI/NheI double digestion and sequencing showed that the target fragment was 2 459 bp and consistent with the corresponding sequence of GenBank; Using the 2459 bp fragment as a template, 5 pairs of primers with identical 3' terminus and different 5' terminus were designed and synthesized for half-nest PCR amplification. 5 fragments with an interval of approximate 400 bp each other, i.e. 2112 bp, 1703 bp, 1290 bp, 784 bp and 496 bp, were produced and inserted into pGEM-T after recovery and purification for transformation to TOP10 competent E. coli. and screening of positive colonies. After that, KpnI/NheI was used to digest the above-mentioned five pGEM-T recombinant plasmids and pGL3 basic vector, and the ligation was completed by T4 DNA ligase. After transformation to TOP10 competent E. coli. and screening for positive colonies, 5 subcloned recombinants of ID4 gene promoter and pGL3 Basic vector cells were constructed. In conclusion, 2.5 kb ID4 gene promoter with upstream expression regulatory sequence was successfully cloned and a series of ID4 promoter subclone-pGL3-Basic recombinant were constructed for further researches on activity, expression regulation and function of ID4 promoter.</p>

Bioinformatics scan of factors with inhibitory effect on lrp16 gene expression / 中国实验血液学杂志

The main purpose of the this study was to find the candidate cis-elements in negative regulation region throngh analysing the DNA sequences of lrp16 gene promoter so as to provide the experimental basis for screening drugs with inhibitory effect on lrp16 gene expression. The open reading frame (ORF) sequences in uncoding DNA and mRNA sequences of 5' flanking region in lrp16 gene were cloned by the data in GeneBank and Internet; the possibly existing cis-element in thsi region was searched in databank of human transcriptional factor by using TESS and Genomax online promoter analysis software; the drugs related to inhibition of lrp16 gene expression were screened by using SAGE and GEO databank. The results showed that there were many cis-elements in the negative regulation region, including T-Ag, PU.1, c-Ets, XPF-1, P2 alphaA, IL6-6RE and RAR. In cultured cell lines, hormone or its inhibitor such as corticosteroid, tamoxifen, forskolin, phenylephrine, inflammatory factors such as IFNgamma and TNFalpha, and chemotherapeutics 5-fluorouracil could down-regulate the lrp16 gene expression as compared with absent ones. It is concluded that cis-elements including T-Ag, PU.1, c-Ets, XPF-1, P2 alphaA, IL6-6RE and RAR may inhibit lrp16 expression and hormone or its inhibitor such as corticosteroid, tamoxifen, forskolin, phenylephrine, inflammatory factors such as IL6, IFNgamma and TNFalpha, and chemotherapeutics 5-fluorouracil may participate in the regulation of lrp16 gene expression in negative manner.

Studies on chemical constituents of Ranunculus japonicus / 中国中药杂志

<p><b>OBJECTIVE</b>To isolate and determine the chemical constituents of Ranunculus japonicus in Liupan Mountain, Ningxia province, China.</p><p><b>METHOD</b>The herb was extracted with ethanol by ultrasonic bath. The extractives were divided to petroleum ether, ethyl acetate, n-butanol parts. The first two parts were separated and purified with silica gel and Sephadex LH -20 column chromatography. The structures of the separated compounds were idnetefied by physical and chemical properties and spectral analysis.</p><p><b>RESULT</b>Nine compounds were isolated and identified as follows: scoparone (1), tricin (2), protocatechuic acid (3), luteolin (4), anemonin (5), scopoletin (6), 5-hydroxy-6, 7-dimethoxyflavone (7), ternatolide (8), 5-hydroxy-7, 8-dimethoxy-flavone (9).</p><p><b>CONCLUSION</b>Compounds 1-9 were isolated from Ranunculus japonicus for the first time.</p>