Research progress and application prospect of stem cells in inflamed dental tissues on periodontal bone regeneration / 中华口腔医学杂志

Tissue engineering has become a research hotspot regarding periodontal bone regeneration in recent years. Generally, stem cells used in periodontal tissue engineering are derived from healthy dental tissues, while restricted due to the strict indication of tooth extraction and limited sources. Stem cells in inflamed dental tissues mainly derive from inflamed pulp, periapical and periodontal tissues. Stem cells in inflamed dental tissues are abundant and retain most of the basic characteristics of stem cell compared with the ones derived from healthy dental tissues, which can be a promising source of stem cells for periodontal bone regeneration. In this review, we summarize the current application and prospect of stem cells in inflamed dental tissues on periodontal bone regeneration, and then discuss their feasibility as seed cells, in order to provide a reference for future research and clinical application of stem cells in inflamed dental tissues.

Efficacy and safety of intratracheal administration of budesonide combined with pulmonary surfactant in preventing bronchopulmonary dysplasia: a prospective randomized controlled trial / 中国当代儿科杂志

OBJECTIVES@#To study the efficacy and safety of early intratracheal administration of budesonide combined with pulmonary surfactant (PS) in preventing bronchopulmonary dysplasia (BPD).@*METHODS@#A prospective randomized controlled trial was designed. A total of 122 infants with a high risk of BPD who were admitted to the neonatal intensive care unit of the Third Affiliated Hospital of Zhengzhou University from January to July 2021 were enrolled. The infants were randomly divided into a conventional treatment group with 62 infants (treated with PS alone at an initial dose of 200 mg/kg, followed by a dose of 100 mg/kg according to the condition of the infant) and an observation group with 60 infants (treated with PS at the same dose as the conventional treatment group, with the addition of budesonide 0.25 mg/kg for intratracheal instillation at each time of PS application). The two groups were compared in terms of the times of PS use, ventilator parameters at different time points, oxygen inhalation, incidence rate and severity of BPD, incidence rate of complications, and tidal breathing pulmonary function at the corrected gestational age of 40 weeks.@*RESULTS@#Compared with the conventional treatment group, the observation group had a significantly lower proportion of infants using PS for two or three times (P<0.05). Compared with the conventional treatment group, the observation group had a significantly lower fraction of inspired oxygen at 24 and 48 hours and 3, 7, and 21 days after administration, significantly shorter durations of invasive ventilation, noninvasive ventilation, ventilator application, and oxygen therapy, a significantly lower incidence rate of BPD, and a significantly lower severity of BPD (P<0.05). There was no significant difference in the incidence rate of glucocorticoid-related complications between the two groups (P>0.05).@*CONCLUSIONS@#Compared with PS use alone in preterm infants with a high risk of BPD, budesonide combined with PS can reduce repeated use of PS, lower ventilator parameters, shorten the duration of respiratory support, and reduce the incidence rate and severity of BPD, without increasing the incidence rate of glucocorticoid-related complications.

Study of electrophysiological mechanism of dopamine inhibiting insulin secretion by Kv channels / 中国药理学通报

Aim To study the electrophysiological mechanism of dopamine inhibiting insulin secretion hv voltage-dependent potassium ( Kv) channels.Methods Islets and (3 cells were isolated from male SD rats.D,-like receptor agonist ( SKP38393), D2-like receptor agonist (Quinpirole) and antagonist (Epiclopride) were used according to the experiment.Insulin secretion was detected by insulin radioimmunoassay.Whole-cell j J patch-clamp technique was applied to detect Kv channel currents and action potential duration of p cells.Di- BAC4(3) staining was used to observe membrane potential.Results SKF38393 did not affect insulin secretion and the Kv channel currents.Quinpirole signifi cantly inhibited insulin secretion and increased Kv channel currents.Dopamine significantly inhibited insulin secretion, increased Kv channel currents and shortened action potential duration of p cells, which could be reversed by epiclopride.In addition, dopamine de-creased membrane potential of INS-1 cells.Conclusions Dopamine inhibits insulin secretion by acting on D2-like receptors, resulting in actived Kv channels, shortened action potential duration and decreased cell membrane potential.

Analysis of codon usage patterns in Polyporus umbellatus based on transcriptome data / 药学学报

We obtained 332 coding sequences from the Polyporus umbellatus transcriptome based on the BLASTx and ESTScan analyses. The codon usage patterns of P. umbellatus were calculated and statistically analyzed using CodonW. The results showed that the average GC content of genes was 53.57% and the average GC3 content was 57.98%, suggesting that genes favored codons ending with G or C. The effective number of codons (ENC) value range from 38.46 to 61, which indicates that these genes have low codon usage bias. The neutrality plot and ENC-plot analysis revealed that many factors such as mutation and selective pressure play an important role in shaping codon usage bias in P. umbellatus genes. Twenty-two optimal codons were identified as being biased toward codons ending with G or C using the high expression superior codon analysis method. This study will lay a foundation for future research on genetic engineering and molecular evolution in P. umbellatus.

The prognostic significance of dynamic monitoring of minimal residual disease (MRD) status in patients with newly-diagnosed multiple myeloma / 中华血液学杂志

Objective: To evaluate the prognostic value of kinetic changes in minimal residual disease (MRD) status, as well as its relationship with risk stratification, therapeutic response and treatment in patients with newly-diagnosed multiple myeloma (MM) . Methods: A total of 135 patients with newly-diagnosed MM were screened, and 105 patients who achieved VGPR or more as the best responses were included into this study. The MRD status was determined by multiparameter flow cytometry (MFC) at multiple intervals after two cycles of treatment until clinical relapse, death, or last follow-up. The statistical methods included Kaplan-Meier analysis, Cox regression, etc. Results: ①In all 135 patients, 57.8% (78/135) patients achieved MRD negativity (MRD(-)) after treatment. In 105 patients who achieved VGPR and thus included in this study, the MRD(-) rate was 72.4% (76/105) , with a median interval of 3 months from starting treatment to achievement of MRD(-) status. ②The 2-year PFS rate of patients with MRD(-) status was significantly higher than that of MRD(+) status (62.2% vs 41.3%, P=0.001) , while MRD persistence (MRD(+)) was an independent factor for poor prognosis (multivariate analysis for PFS: P=0.044, HR=3.039, 95%CI 1.029-8.974) . ③Loss of MRD(-) status (i.e., MRD reappearance) showed inferior outcomes compared with MRD sustained negative ones, the PFS was 18 months versus not reach (P<0.001) and the OS was not reach for both (P=0.002) . ④The 2-year PFS and OS rates of patients with duration of MRD(-)status≥12 months were significantly higher than those of the control group (PFS: 77.7% vs 36.7%, P<0.001; OS: 96.4% vs 57.9%, P<0.001 respectively) . Duration of MRD(-) status was associated with a marked reduction in risk of relapse or death (univariate analysis for PFS: P<0.001, HR=0.865, 95%CI 0.815-0.918; for OS: P=0.001, HR=0.850, 95%CI 0.741-0.915 respectively) . ⑤Moreover, even in patients carrying high-risk cytogenetic abnormalities (CA) or ineligible for ASCT, MRD negativity remained its prognostic value to predict PFS (high-risk CA medianPFS: not reach vs 19 months, P=0.006; ineligible for ASCT medianPFS: not reach vs 25 months, P=0.052 respectively) . ⑥Last, treatment with the bortezomib-based regimens contributed to prolonged MRD(-) duration (median MRD(-) duratio: 25 months vs 10 months, P=0.034) . Conclusion: Our findings supported MRD(+) status as an independent poor prognostic factor in MM patients, which implicated that duration of MRD(-) status also played a significant role in evaluation of prognosis, while loss of MRD(-)status might serve as an early biomarker for relapse. Therefore, monitoring of MRD kinetics might more precisely predict prognosis, as well as guide treatment decision, especially for when to start retreatment in relapsed patients.

High-throughput transcriptomic sequencing of Rheum palmatum L. seedlings and elucidation of genes in anthraquinone biosynthesis / 药学学报

Anthraquinones are not only the main active constituents but also the index components for the quality control of Rhei Radix et Rhizoma. To study the anthraquinone biosynthesis, Rheum palmatum L. seedlings were subjected to a high-throughput transcriptomic sequencing analysis by Illumina HiSeqTM 2000 150PE. The Illumina sequencing generated a total of 11.04 G clean data resulting in 736 309 74 clean reads, deposited in the sequence read archive (SRA accession SRP160030). Trinity do novo assembly yielded 93 646 unigenes, with an average of 1 108 nt. Functional annotation revealed that all unigenes were successfully annotated in the NR, NT, Swiss-port, PFAM, and KOG databases. GO enrichments showed that 57 subgroups were involved in biological process, cellular component, and molecular function. KEGG analysis indicated that 1 107 unigenes were implicated in 19 standard secondary metabolic pathways. 172 unigenes were analyzed to encode 28 key enzymes during the MVA, MEP, shikimic acid, and polyketide pathways related to anthraquinone biosynthesis. 125 CYP450 and 73 UGTs unigenes were related the modification of secondary metabolites in R. palmatum L. Furthermore, seven unigenes with full length cDNAs were successfully verified by RT-PCR and sequencing analyses. Then, MISA prediction produced a number of 18 885 simple sequence repeats (SSRs). Herein, the transcriptomic gene expression profiles of R. palmatum L. and candidate genes during the anthraquinone biosynthesis pathway were obtained for the first time. The results provided basic information for subsequent gene function characterization, secondary metabolic pathway analysis, and anthraquinone biosynthesis and regulation elucidation in R. palmatum L.

Effect of baicalein on pancreas insulin secretion in rats and its mechanism / 中国药理学通报

Aim To investigate the effect of baicalein on insulin secretion from rat islets and the underlying mechanism. Methods Pancreatic islets were obtained from the pancreas of male Wistar rats by collagenase P digestion and histopaque-1077 density gradient separa-tion. Single islet cells were dispersed from pancreatic islets by Dispase II digestion. Insulin secretion experi-ment was applied to observe insulin release after baica-lein stimulation. To study the potential mechanism, calcium imaging technique and patch-clamp experiment were applied to measure intracellular Ca2+ concentra-tion and voltage-dependent potassium channel currents (Kv). Results In 16. 7mmol·L-1 glucose, baica-lein accelerated insulin secretion in a dose-dependent manner. Baicalein promoted the intracellular Ca2+ con-centration. The patch-clamp experiment showed that baicalein inhibited Kv current in a dose-dependent manner. Conclusion Baicalein can increase the in-tracellular Ca2+ concentration by inhibiting Kv chan-nels and eventually promoting insulin secretion.

Effects of estrogen receptor antagonist on the expressions of Erα, Erβ and p57kip2 in JEC cells of human endometrial carcinoma / 临床与实验病理学杂志

Purpose To explore the effects of estrogen receptor antagonist on the expression of estrogen receptor subtype (ERα, ERβ), and p57kip2 protein in human endometrioid carcinoma cells named JEC. Methods The JEC cells (moderately differentiated EC cells) cultured in vitro were treated with β-Estradiol (E2) (10～6 mol/L) and two types of estrogen receptor antagonists, tamoxifen (TAM) and fulvestrant (ICI182780) (10-6 mol/L). After 24, 48, 72 h, MTT was used to detect the growth condition of JEC cells, and the light microscopy and electron microscopy were used to observe the growth condition and morphological changes of cells, Western blot was used to detect the expression of ERα, ERβ, PR-A, PR-B and P57kip2 protein in JEC cells. Results MTT results: Compared with the control group, E2 could promote the proliferation of JEC cells significantly (P＜0.05), and ICI182780 could inhibit the proliferation of JEC cells obviously (P＜0.05). Compared with the E2 group, the proliferation ability of JEC cells in E2 + ICI182780 group were lower(P＜0.05). Morphological change: Compared with the control group, the cells density of E2 group increased obviously, and the pathologic mitosis was easy to seen in some cells. The cells density decreased obviously in ICI182780 group. Compared with E2 group, the cells density of E2 + TAM group and E2 + ICI182780 group were decreased, and pathological mitotic figures were difficult to seen. Western blot results: Compared with the control group, the expression of ERβ protein increased, and the expression of p57kip2 protein decreased in E2 group (P＜0.05). The expression of ERβ protein decreased, and the expression of p57kip2 protein increased in ICI182780 group and TAM group, and the difference was statistically significant between ICI182780 group and control group (P＜0.05). Compared with the E2 group, the expression of ERβ protein decreased, and the expression of p57kip2 protein increased in E2 + ICI182780 group and E2 + TAM group, and the difference was statistically significant between E2 + ICI182780 group and E2 group (P＜0.05). ERa protein of JEC cells did not expressed in experimental group or control group. Conclusion ERa protein are not expressed in JEC cells. ICI182780 have a stronger role in antagonizing estrogen, and may induce the expression of p57kip2 protein by down-regulating the expression of ERβ protein in JEC cells, block the cell cycle progression and inhibit the growth of tumor cells. TAM has a weaker estrogen like effect on the growth of JEC cells. It is possible that combined detection of the expression of ERa and p57kip2 protein in EC has an important reference value for individualized selection of endocrine therapy for EC patients.

Expression of SEL1L and BCL-2 in diffuse large B cell lymphoma / 临床与实验病理学杂志

Purpose To investigate the expression and the clinical significances of SEL1L and BCL-2 in 123 cases of diffuse large B cell lymphoma (DLBCL) and cell line SUDHL-4, LY-10. Methods Immunohistochemistry staining for SEL1L was performed in 123 DLBCL and 60 reactive lymphoid hyperplasia (RLH), and also BCL-2 protein in 123 DLBCL. Immunocytochemistry staining and Western blot analysis for SEL1L protein were used in SUDHL-4 and LY-10. Results The high expression rate of SEL1L was 69.9％ in 123 DLBCL, which was significantly higher than that in 60 RLH (25.0％ ). The expression of SEL1L protein in DLBCL was not related to clinic pathological parameters. The positive rate of BCL-2 was 83.7％ in123 DLBCL. The expression of BCL-2 protein was correlated with immunophenotyping, primary location, and Ann Arbor stage. The expression of SEL1L protein was positively correlated with that of BCL-2 protein in DLBCL (r=0.227, P＜0.05). SEL1L protein was also detected in SUDHL-4 and LY-10 cell lines. Conclusion The SEL1L protein may play an important role in the carcinogenesis of DLBCL, and may be associated with BCL-2.

Molecular cloning and prokaryotic expression of a type II ribosome inactivating protein from Polyporus umbellatus / 药学学报

A type Ⅱ ribosome inactivating protein (RIP) gene was cloned from Polyporus umbellatus sclerotia by RT-PCR method. The full open reading frame cDNA sequence of this gene was 873 bp in length and encoded a 290-aa protein with a molecular weight of 32.33 kDa and an isoelectric point of 5.58. Multiple sequence alignment revealed that the deduced amino acids possessed conserved domains of RICIN superfamily protein. A neighbor joining phylogenetic analysis suggests that PuRIP was closely related to RIP in Marasmius oreades. Real time PCR results showed that this gene expressed in all tested tissues of P. umbellatus. Meanwhile, the expression of this gene was significantly up-regulated in the part infected by Armillaria mellea. This result suggested that this PuRIP might played important role with potential biotic stress tolerance of P. umbellatus. Otherwise, we successfully constructed the pET15b-PuRIP plasmid, produced and purified the His-PuRIP fusion protein, which would provide the basic material for polyclonal antibody preparation and gene function research.

Cloning and Expression Analysis of Nine Major Facilitator Superfamily Encoding Genes in Polyporus umbellatus / 中国药学杂志

OBJECTIVE: To clone and isolate the major facilitator superfamily(MFS)genes of Polyporus umbellatusand carry out bioinformatic analysis. METHODS: Nine major facilitator superfamily(MFS)genes were cloned fromPolyporus umbellatus sclerotia by RT-PCR and the expression analysis of the nine genes in different parts ofPolyporus umbellatus sclerotia was carried out using quantitative Real-time PCR.RESULTS: The full open reading frame cDNA sequence of these nine genes was between 1 321 and 1 860 bp, the putative encoding proteins were between 441 and 620 amino acids, the molecular weight was between 48.45×103 and 64.79×103 and the theoretical pI was between 6.59 and 9.56. The amino acids of these nine genes possessed 11 to 14 membrane-spanning domains. Phylogenetic tree analysis indicated that Comp34750, Comp34832, Comp29252, Comp42895, Comp32579 and Comp27555 had the highest similarity with MFS general substrate transporter, Comp28872 andComp26306 had the highest similarity with MFS monosaccharide transporter, and Comp33117 had the highest similarity with MFS sugar transporter. Quantitative real-time PCR showed that these nine genes were expressed in both the symbiotic part and non-symbiotic part. Meanwhile, the expressions of seven genes were significantly up-regulated in the symbiotic part except Comp34382 and Comp32579. CONCLUSION: The investigated nine genes might play an important role during the defense response and nutrient absorption of P.umbellatus.

Cloning, Sequence Analysis and Expression Analysis of Eleven Cytochrome P450 Encoding Genes in Polyporus umbellatus / 中国药学杂志

OBJECTIVE: To clone the cytochrome P450 genes from a medicinal fungus Polyporus umbellatus and carry out bioinformatic analysis. METHODS: The full length cDNAs of the genes were obtained by RT-PCR. Some bioinformatic tools were used to characterize the physiochemical properties of the 11 deduced protein. The analyses of multiple alignment and phylogenetic trees were performed with MEGA 6.0 software. RESULTS: Eleven P450 genes were cloned using RT-PCR method from the Polyporus umbellatus sclerotium. The full open reading frame cDNA sequence of these eleven genes was between 1 326 and 1 635 bp, the putative encoding proteins were between 442 and 545 amino acids, the molecular weight was between 48.9×103 and 61.5×103, and the theoretical pI was between 6.3 and 8.9. Protein sequence analysis found that there was conserved P450 domain in the 11 genes. Phylogenetic tree analysis indicated that all these 11 P450 genes belonged to basidiomycetes. Quantitative real-time PCR showed that these 11 genes were expressed in both the infected partand non-infectedpart. Meanwhile, the expressions of seven genes were significantly up-regulated in the infected part except comp18720, comp26906, comp32003 or comp33717. CONCLUSION: Molecular characterization of the 11Cytochrome P450 genes will be useful for further functional determination of the genes involved in the defense progress of Polyporus umbellatus.

Molecular cloning and characterization of an inorganic phosphate transporter protein encoding gene in Polyporus umbellatus / 中草药

Objective To clone an inorganic phosphate transporter (PiT) gene from Polyporus umbellatus and perform the bioinformatics and expression mode analysis. Methods Using RT-PCR.to clone the full-length cDNA of PiT. The characteristics of physiochemical properties, conserved domains, signal peptide and transmembrane domain of the predicted PiT protein were determined by using bioinformatic tools. Results A inorganic phosphate transporter (PiT) gene (NCBI: KU179154), designated as PuPiT, was cloned from Polyporus umbellatus sclerotia by RT-PCR. The full open reading frame cDNA sequence of PuPiT was 1 590 bp, encoding a putative PiT protein with 530 amino acids with a molecular weight of 57 552, and a theoretical pI of 6.82. The amino acids possess 12 membrane-spanning domains. Phylogenetic tree analysis indicated that PuPiT had the highest similarity with PiT from Moniliophthora rorer, and had high similarity with Moniliophthora roreri, Laccaria bicolor, and Heterobasidion irregulare. Quantitative real-time PCR showed that PuPiT expressed in both the symbiotic part and non-symbiotic part. Meanwhile, the expression of PuPiT in the symbiotic part was significantly up-regulated, about 12 times more than that in the non-symbiotic part. This result showed that PuPiT might play an important role in the Pi accumulating. Conclusion Molecular cloning and characterization of the novel PuPiT gene will be useful for further functional determination of the gene involving in phosphorus translocation regulation and symbiotic process.

Molecular cloning and characterization of two kind of heat-shock protein gene from Polyporus umbellatus / 中国中药杂志

With RT-PCR approaches, the full-length cDNA of two heat shock protein genes were cloned from total RNA of the Polyporus umbellatus sclerotium. The full open reading frame cDNA sequence of the Hsp90 was 2 091 bp, encoding 696 amino acid residues with a predicted molecular mass of 78.9 kDa. The full open reading frame cDNA sequence of the Hsp70 was 1 944 bp, encoding 647 amino acid residues with a predicted molecular mass of 70.5 kDa. The Hsp90 and Hsp70 protein contained the conservative structure domain, respectively. Phylogenetic analysis showed that Hsp90 and Hsp90 from Trametes versicolor were clustered into one group, Hsp70 and Hsp70 from Fistulina hepatica were clustered into one group. Real-time PCR analysis showed that, the expression of Hsp90 and Hsp70 in the infected part by Amillariella mellea was upregulated. The expression profiling of Hsp90 and Hsp70 showed same patterns underbiotic stress. The results indicate that these two genes may play an important role in response to Amillariella mellea infection.

Habitat suitability assessment of medicinal Polyporus umbellatusin China based on Maxent modeling / 中国中药杂志

Geographic distribution of Polyporus umbellatus was predicted by using distribution records. Based on 42 distribution records from 12 provinces and bioclimatic data (1950-2000), georaphic distribution of P. umbellatus was modeled using Maxent. The results showed thatthe Receiver Operating Characteristic (ROC) curve analysis method was used to assess the accuracy of MAXENT model and the area under ROC curve (AUC) value of MAXENT was 0. 960 which suggested that the result of assessment was dependable. The geographic distribution pattern of were divided into three distribution block based on distribution values of 0.5-0.8: small area of Heilongjiang, Jilin, Liaoning and Hebei province, the board area of Yunnan, Guizhou and Sichuan, the southeast area of Tibet and the most area of Shanxi and Shannxi, the southeast board area of Shannxi, Gansu and Ningxia. Jackknife Test showed that average precipitation in warm seasons had the greatest contribution to the distribution gain of P. umbellatus, followed by mean temperature of driest quarter and annual mean temperature. The object suggests the potential distribution areasof P. umbellatus which is useful for the habitat conservation and introduction of P. umbellatus.

Clinical Research about Risk Factors and Reversible Predictors in Renal Impairment due to Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was aimed to investigate the risk factors of renal impairment and the predictive factors of renal function recovery so as to provide basis for its prevention and treatment.</p><p><b>METHODS</b>Medical records of 161 patients with MM firstly diagnosed from January 2007 to April 2013 were analyzed retrospectively. Among them 58 cases accompanied with renal insufficiency (group A, others belong to group B) and 39 of them regain normal renal function after some treatment. The possible related renal impairment risk factors and reversible predictors were analyzed with chi-square test for significance firstly, then factors that have significant difference were entered into multivariate logistic regression analysis.</p><p><b>RESULTS</b>Systolic blood pressure (SBP), hemoglobin, uric acid, blood calcium, phosphorus, serum β2-microglobulin, urine β2-microglobulin levels, M-component type, light chain type, nephrotoxic drug use, infection in group A had significant difference (P<0.05) compared with those in group B; the systolic blood pressure, diastolic blood pressure, platelet, globulin, blood calcium, and urine β2-microglobulin levels, the chemotherapy applied and the response to chemotherapy in reversed group were significantly different from no-reversed group (P<0.05). Multivariate logistic regression showed that light chain type, Hb, uric acid, Ca were the independent risk factors for the development of renal failure in MM, and Ca, chemotherapy and the response to chemotherapy were the predictors of renal function recovery.</p><p><b>CONCLUSION</b>High blood calcium, severe anemia, λ light chain, high uric acid are the independent risk factors of renal impairment in MM patients. Patients with high blood calcium before treatment easily regain normal renal function after effective chemotherapy. Bortezomib-based chemotherapy has higher response rate and higher reversal rate, and it may be related with its unique mechanism.</p>

Investigation of symbiotic Armillaria species with Chinese traditional medicinal fungus Polyporus umbellatus / 中国药学杂志

OBJECTIVE: To identify symbiotic Armillaria species with Polyporus umbellatus. METHODS: Armillaria was cultured on PDA paltes. To determine whether the strains were pure culture, the internal intergenic spacer (IGS) region were sequenced. Furthermore, the NCBI BLAST program was used to search similar sequences in the GenBanksequence database for the IGS sequence of the species on homology, and a phylogenetic tree was constructed based on neighbor-joining method. RESULTS: All isolates were similar in colony morphology and grew well in PDA medium with well-developed rhizomorphs. The isolated 22 strains were pure culture of the fungus. Phylogenetic analysis based on IGS sequence indicated that the symbiotic Armillaria species with Polyporus umbellatus belonged to A. cepistipes, A. gallica, A. ostoyae, and A. mellea. CONCLUSION: In the present study, phylogenetic analysis based on molecular method was carried out for the symbiotic Armillaria species with P. umbellatus for the first time. The results suggest that Armillaria species have no specificity with P. umbellatus, which is different from the previous reports. Further research will be done on the growth of P. umbellatus affected by different kinds of Armillaria species.

Cloning and expression analysis of Bax inhibitor-1 gene from medicinal fungus Polyporus umbellatus / 中草药

Objective: To clone the Bax inhibitor-1 (BI-1) gene from Polyporus umbellatus and perform the bioinformatics and expression mode analysis. Methods: To clone the full-length cDNA of BI-1 using RACE technology. The characteristics of physiochemical properties, conserved domains, and transmembrane domain of the predicted BI-1 protein were determined using bioinformatic tools. Results: The full-length cDNA of BI-1 gene was 1 091 bp in length and encoded a 334-aa protein with a molecular weight of 36170 and an isoelectric point (pI) of 10.51. The polypeptide chain was a hydrophobin with five hydrophobic regions. The PuBI-1 belonged to basidiomycete group according to the phylogenetic analysis. Real time quantitative PCR (RT-qPCR) analysis revealed that the transcription level of PuBI-1 gene was significantly higher in the beginning and developing stages of sclerotial formation with 3.8 and 7.497 fold over those in the mycelium, but the transcripts decreased sharply with the sclerotial development. Conclusion: Molecular characterization and expression patten of PuBI-1 gene will be useful for the further functional determination of the gene during the development of P. umbellatus sclerotium.

Molecular cloning and characterization of four small GTPase genes from medicinal fungus Polyporus umbellatus / 药学学报

Four small GTPase genes which may be relative to sclerotial development were firstly cloned from medicinal fungus Polyporus umbellatus using rapid amplification of cDNA end PCR (RACE) method. The results showed that full-length cDNA of PuRhoA was 698 bp contained 585 bp ORF, which was predicted to encode a 194 amino acid protein with a molecular weight of 21.75 kD with an isoelectric point (pI) of 6.44; the full length cDNA of PuRhoA2 was 837 bp in length and encoded a 194 amino acid protein with a molecular weight of 21.75 kD and an isoelectric point (pI) of 6.33; the full length cDNA of Puypt1 was 896 bp in length and encoded a 204-aa protein with a molecular weight of 22.556 kD and an isoelectric point (pI) of 5.75; the full length cDNA of PuRas was 803 bp in length and encoded a 212-aa protein with a molecular weight of 23.821 kD and an isoelectric point (pI) of 5.2. There are fani acyl transferase enzyme catalytic site and myrcene-transferase enzyme catalytic site in PuRhoA1 while the PuRhoA2 only possess myrcene-transferase enzyme catalytic site. Puypt1 contains the Rab1-Ypt1 conserved domain of small GTPase family and PuRas contains the fani acyl transferase enzyme catalytic site. According to the phylogenetic analysis all these four small GTPase clustered with basidiomycete group. Quantitative real-time PCR analysis revealed that Puypt1, PuRas and PuRhoA1 transcripts were significantly higher in the beginning of sclerotial formation than that in the mycelia, whereas the transcripts levels of PuRhoA2 gene were particularly lower in sclerotia than that in mycelia, suggesting that these four genes might be involved in P umbellatus selerotial development.

Expressive changes of CD4(+)T cell subset transcription factors in patients with aplastic anemia, myelodysplastic syndrome and acute myeloid leukemia and their clinical significances / 中国实验血液学杂志

This study was aimed to compare the expressions of specific transcription factors of CD4(+) T cell subset ( T-bet, GATA-3, RORγt and FoxP3 mRNA) in peripheral blood of patients with aplastic anemia(AA), myelodysplastic syndrome(MDS), and acute myeloid leukemia(AML), and investigate their immune status and pathogenesis, so as to provide experimental basis for the choice of clinical treatment. The expression of T-box (T-bet), GATA-3, ROR-γt and Foxp3 mRNA in PBMNC were examined by RT-PCR in 42 cases of MDS, including 22 refractory anemia(MDS-RA) and 20 refractory anemia with excess blasts (MDS-RAEB), in 23 cases of AA, 17 cases of AML patients and 16 healthy volunteers respectively. The results indicated that, compared with normal control group, expressions of T-bet and RORγt mRNA in AA patient group were significantly higher (P < 0.01), expression levels of GATA3 Foxp3 mRNA were lower (both P < 0.01). There was no significant difference in expression of T-bet and GATA3 mRNA between MDS group and normal control group, but the expression levels of Foxp3 and RORγt mRNA were higher than those in normal controls (P < 0.05); T-bet and RORγt in MDS-RA group were higher than those in the normal controls(P < 0.01), and GATA3 expression significantly reduced (P < 0.05), however, there was no significant difference in expression of Foxp3 between MDS-RA and the controls. Expression levels of T-bet and RORγt mRNA in patients with MDS-RAEB and AML were lower than those in normal controls (P < 0.05), but the expression levels of GATA3 and Foxp3 mRNA were significantly higher than those in normal controls (P < 0.01). It is concluded that the transcription factor expressions are different in PBMNC of patients among these three diseases. Immune-mediated excessive apoptosis may play an important role in pathogenesis, bone marrow failure in patients with AA and MDS-RA, and abnormal clones of immature cells may be one of main reasons for bone marrow failure in AML and late stage of MDS.