The Expression and Clinical Correlation of the CD68 Factor in Acute Myeloid Leukemia Patients / 中国实验血液学杂志

OBJECTIVE@#To analyze the expression and clinical characteristics of CD68 in bone marrow and peripheral blood of patients with acute myeloid leukemia (AML).@*METHODS@#The expression of CD68 in bone marrow blast cells was detected by four-color flow cytometry in 50 newly diagnosed AML patients and 23 controls. The expression of CD68 in peripheral blood of 85 newly diagnosed AML patients, 29 remission AML patients and 24 controls was detected by ELISA. The correlation between the expression rate of non-M3 AML bone marrow CD68, peripheral blood CD68 concentration and white blood cell count and other clinical data was compared respectively.@*RESULTS@#The median CD68 expression rate in myeloid leukemia cells of non-M3 AML patients was 19.7%, significantly higher than control (0.2%) (P<0.001). The median concentration of non-M3 CD68 in peripheral blood was 67.97 pg/ml, significantly higher than in control (29.94 pg/ml)（P<0.01）. There was no statistically significant difference in the plasma CD68 concentration of the peripheral blood between the newly diagnosed (45.72 pg/ml) and the remission stage (55.12 pg/ml) of non-M3 AML patients by paired analysis (P>0.05). The results showed that the higher the expression rate of CD68 in bone marrow, the higher the count of white blood cells in peripheral blood, and the lower the count of hemoglobin and platelet in peripheral blood. The higher the plasma concentration of CD68 in peripheral blood, the higher the white blood cell count and the lower the complete remission rate.@*CONCLUSION@#The expression of CD68 both in bone marrow and peripheral blood of patients with non-M3 AML is higher than that of control group. Patients with high expression of CD68 show a low rate of complete remission, suggesting that the expression level of CD68 is correlated with treatment response.

Establishment of Acquired Aplastic Anemia SD Rat Model / 中国实验血液学杂志

OBJECTIVE@#To establish a stable and rapidly rat model acquired aplastic anemia.@*METHODS@#The SD rats were exposed to Csγ-ray at 3.5 and 4.0 Gy ( 91 cGy/min), and intraperitoneally injected with CTX at 35 mg/( kg·d) and CHL at 45 mg/( kg·d) in d 4, 6 and 8 after irradiation; the WBC, platelet and reticulocyte counts in peripheral blood, the smears and nucleated cells counts of bone marrow were observed.@*RESULTS@#The levels of peripheral blood 3-lineage cells of SD rats treated with Csγ-irradiation combined with cyclophosphamide and chloramphenicol were significantly reduced, among which white blood cells, platelets and reticulocytes decreased rapidly, and the number of bone marrow nucleated cells decreased significantly; bone marrow pathological sections showed severe reduction of hematopoietic cells, and the non-hematopoietic cells such as fat cells increased, and a serve or lightly reduction of bone marrow cells were found.@*CONCLUSION@#The rat model established by Csγ-ray irradiation combined with cyclophosphamide and chloramphenicol meets the clinical characteristics of aplastic anemia, and this study provides a stable rat model for the study of new therapeutic drugs for acquired aplastic anemia.

Establishment of Cs γ ray Combined with Cyclophosphamide and Chloramphenicol-Mediated Mouse Model of Acquired Aplastic Anemia / 中国实验血液学杂志

OBJECTIVE@#To improve and establish the mouse model with aplastic anemia (AA) mediated by Cs γ-ray irradiation combined with cyclophosphamide (CTX) and chloramphenicol (CHL) injection,so as to provide a stable model for studying the pathogenesis and treatment of AA .@*METHODS@#The BALB/c mice were exposed to Cs γ-ray of 3-5 Gy(91 cGy/min) and were intraperitoneally injected with CTX of 25 mg/(kg.d) and CHL of 62.5 mg/(kg.d) at D 4,5 and 6 after irradiation; the WBC, platelet and reticulocyte counts in peripheral blood as well as the mucleated cell count in bone marrow and bone marrow smears were detected .@*RESULTS@#The 3-lineage cells in peripheral blood of BALB/c mouse model with acquired AA were rapidly reduced, especially WBC, platelet and reticulocyte counts were lowest at D 14,the 3-lineage cells in peripheral blood were still severely reduced at D 28; the nucleated cell count in bone marrow significantly dcreased,the bone marrow hyperplasia was reduced or severely reduced; the pathological sections of bone marrow showed the severe reduction of hematopoietic cells and the increased of non-hematopoietic cells such as fat cells.@*CONCLUSION@#The mouse model with acquired AA has been established by Cs γ-ray irradiation combined with CTX and CHL injection. All detection indicators of this model reach to diagnostic criteria for acquired AA,therefore this mouse model may be used as the model for study of pathogenesis and treatment of acquired AA.

Mycorrhizal Diversity Associated to Liparis japonica (Miq.) Maxim. in China

Orchidaceae (Orchidaceae) is the second largest family of angiosperms. It’s the "flagship" group in plant protection. The existence of orchid plant is closely related to mycorrhizal fungi. The relationship between orchids and their symbiotic mycorrhizal fungi is a benefit to the protection and population restoration of orchids. Aims: The research was aimed to study molecular identification about 15 strains of mycorrhizal fungi from 6 plots by rDNA ITS technology in order to understand and utilise the mycorrhizal fungi of Liparis japonica (Miq.) Maxim. Study Design: The mycorrhizal fungi collected from different geographical locations were isolated and purified from the mycorrhizal fungi symbiotic with Liparis japonica in Northeast China, which were identified by rDNA ITS, meanwhile computed evolutionary distance and constructed the phylogenetic tree. Place and Duration of Study: In 2017, the root segments of Liparis japonica were collected separately from Qianshan, Changbaishan, Gaoguan, Guanmenshan, Dongling, Daqinggou. Methodology: Fifteen strains of mycorrhizal fungi collected from six plots were identified by rDNA ITS. Using DNAMAN software to analyse, the pairwise homology was compared by using the optimal global sequencing option. The evolutionary distances of fifteen strains were calculated by MEGA (Molecular Evolutionary Genetics Analysis) software package and their phylogenetic trees were constructed by neighbour-joining method. Results: With primers ITS1 and ITS4, the 15 mycorrhizal fungi strains of rDNA ITS got about 600 bp length. ITS length was about 582-613 bp, in which ITS1 length was about 177-190 bp, and ITS2 length was 246-273 bp. The mycorrhizal fungi strains were highly homology separated from one plot, mostly above 90%. The plots from the south to the north were as follows: Qianshan, Guanmenshan, Gaoguan, Dongling, Changbaishan, Daqinggou in China. Fifteen strains after separated and purified were identified to be the Epulorhiza of Orchid Rhizoctonia blasted with Genbank. The homology of the strains gradually decreased affected by the difference of the north and the south, namely there was an increasing trend of diversity from south to north. Conclusion: The homology of mycorrhizal fungi from one plot was higher because of the same soil environment and climate environment and so on, and strain type was single. Under the influence of microclimate in Northeast China, the homology of strains decreased gradually in the sample area, that is, the diversity gradually increased from the south to the north.

Meta-analysis on the Safety and Efficacy of Absorb Bioresorbable Vascular Scaffold Versus Everolimus Eluting Stent for Percutaneous Coronary Intervention / 中国循环杂志

Objectives:To systemically review the safety and efficacy of bioresorbable vascular scaffold (BVS) versus everolimus eluting stent (EES) for percutaneous coronary intervention (PCI). Methods:The database searched includes PubMed, Medline, MEDILINE, EMBASE, Cochrane library, CNKI and Wanfang. Database retrieval time was between database establishment time to October 2017. During the same time, authors accessed the conference summary and related websites to collect published randomized controlled trials of published data. To evaluate the quality of the literature according to the modified Jadad scale and extracted the data. Meta-analysis was performed using Review Manager 5.3 software. Results:Nine trials were included; 6 721 patients were randomized to receive BVS (n=3 670) or EES (n=3 051). Time of follow-up was ranged from 6 to 36 months. Compared with metallic EES, risk of target lesion failure (RR=1.31, 95%CI:1.08-1.58; P=0.005) and in-stent thrombosis (RR=2.89, 95%CI:1.85-4.53; P＜0.0001), ischemia-driven target lesion revascularization (RR=1.44,95%CI:1.12-1.86, P=0.005)、target-vessel myocardial infarction (RR=1.74, 95%CI:1.33-2.27, P＜0.0001) and all myocardial infarction (RR=1.49, 95%CI:1.16-1.91, P=0.002) were all significantly higher in BVS group than in EES group. There were no significant differences in all-cause death (RR=0.87, 95 % CI:0.57-1.33, P=0.520), cardiovascular mortality (RR=0.78, 95%CI:0.54-1.11, P=0.160) and composite endpoints (RR=1.10, 95%CI:0.95-1.27, P=0.210) between the two groups. Conclusions:Compared with metallic EES, the BVS appears to be associated with both lower efficacy and higher thrombotic risk during the observation period.

Antidepressant Effect and Mechanism of Honokiol on Acute and Chronic Stress Mouse / 中国药学杂志

OBJECTIVE: To abserve the antidepressant effect and mechanism of honokiol on acute and chronic stress mouse. METHODS: Forced swimming model of acute stress (FST) and chronic stress mice model were used. The acute stress mouse were randomized into a control group, a fluoxetine group (3.3 mg·kg-1), honokiol groups (2.5, 5, 10 mg·kg-1). The chronic stress mouse were randomized into a blank group, a model group, a fluoxetine group(3.3 mg·kg-1), honokiol groups (2.5, 5, 10 mg·kg-1). Then, the immobility time of forced swimming, 5-hydroxytryptamine (5-HT) and 2, 3- indole dioxygenase (IDO) contents in mouse brain tissue by Elisa Kit, and the expression of IDO mRNA in brain tissue used by quantitative real-time PCR were studied. RESULTS: (1)After acute stress, the immobility time of forced swimming in each treatment group was significantly shorter than that in the model group (P<0.05). The 5-HT content of fluoxetine and honokiol medium and high dose group was significantly higher than that of model group (P<0.05, P<0.01). The IDO content of honokiol high dose group was significantly higher than that of model group (P<0.05). (2)After chronic stress, the immobility time of the model group were significantly higher than the blank group (P<0.01). The 5-HT content in brain tissue of the model group was significantly lower than that of the blank group (P<0.01), the IDO content in brain tissue and its expression level of mRNA increased comparing with the normal group (P<0.01). For each treatment group, the immobility time of forced swimming, IDO content and the expression level of IDO mRNA was significantly decreased, and the 5-HT content was significantly increased, comparing with the model group with significant difference (P<0.05). CONCLUSION: Honokiol can relieve the depression behavior of mouse and have certain antidepressant effect. The main mechanism may be associated with the increase of 5-HT, reduction of the tryptophan pathway enzyme IDO content and its gene expression level.

Expression of MicroRNA-148/152 Family in the Hematological Malignancies / 中国实验血液学杂志

MicroRNAs (miRNAs) are small noncoding RNA molecules containing 18-22 nucleotides that regulate gene expression through imperfect interactions with sequences in the 3'-untranslated region (3'-UTR) of target genes. Members of the microRNA-148/152 (miR-148/152) family, which include microRNA-148a (miR-148a), microRNA-148b (miR-148b) and microRNA-152 (miR-152), are of great importance in the development of some tumor diseases. Several studies have demonstrated that members of the miR-148/152 family were expressed differently in many kinds of malignancies, and they play a variety of biological functions, such as regulating tumor growth, proliferation, apoptosis, angiogenesis, and sensitivity to drugs through regulating the expression of target genes. The miR-148/152 family is regulated by methylation of their CPG islands, which reduce the expression of miR-148/152 family members. Interaction has been observed between DNA methylation and miR-148/152 family members through their target gene: DNMT1/3b, an important gene for DNA methylation. So expressions of DNMT1/3b are inversely restricted to the expression level of miR-148a/152. This might result in overexpression of DNMT1/3b, promoting DNA methylation. And then, more DNMT1/3b can be expressed. A novel miR-148a/152-DNMT1/3b regulatory circuit might exist in tumors. Epigenetic abnormalities, especially high methylation of promoter play an important role in occurrence and development of hematological malignancies. Demethylation treatment has become another important way for the treatment. This article summarizes the expression of miR-148/152 family in hematological malignancies, aiming at expounding the signicance of relationship between DNA methylation modification and microRNA.

A study on application of BacT/ALERT 3D liquid culture technology on tuberculosis drug resistance detection / 预防医学

Objective To evaluate the BacT/ALERT 3D liquid culture technology on the detection of drug resistance of Mycobacterium tuberculosis(MTB)and to compare the difference between this technology and Lowenstein -Jensen (L -J) proportion method.Methods BacT/ALERT 3D liquid culture technology and L -J proportion technology were applied to detect the drug resistance of tuberculosis from the positive cultures of 219 solid culture samples.Results The average detection time of BacT/ALERT 3D method was 8.02 ±3.85 d,which was about 20 days shorter than that of L -J proportion method.60 drug resistance strains were found using BacT/ALERT 3D technology,While 79 drug resistance strains were found using L -J proportion technology.There showed no significant difference (P >0.05).The compliance rate of BacT/ALERT 3D method and L -J proportion method on the anti -tuberculosis drugs INH,RFP,EMB and SMwas 95.43%,92.69%,95.43% and 92.24% respectively.Conclusion BacT/ALERT 3D liquid culture technology could detect drug resistant TB strains rapidly with high concordance with the results of L -J proportion method on anti -tuberculosis drugs.

The effect evaluation on the application of BacT/ALERT 3 D technique for the fast cultivation of mycobacterium tuberculosis / 预防医学

Objective To evaluate the BacT/ALERT 3D liquid rapid culture system for the rapid detection of mycobacterium tuberculosis(MTB)and early diagnosis of pulmonary tuberculosis.Methods BacT/ALERT 3D liquid culture mediums and lowenstein-jensen(L-J)solid culture mediums were applied to detect Mycobacterium tuberculosis sputum specimens respectively.Analysis of detection results and detection time were also performed.Results Average positive days of BacT/ALERT 3D liquid culture medium was(13 ±2.05)days,which was shorter than that of L-J solid culture mediums(34.7 ±4.76 )d (P<0.05 ).Compared to the L -J solid culture mediums,BacT/ALERT 3D liquid culture medium had higher positive rate for 59 patients whose sputum smear test was positive,and the positive rate were 71.18%(42/59)and 67.80%(40/59)respectively(P<0.05).BacT/ALERT 3D liquid culture medium had higher positive rate than L-J solid culture mediums for 106 patients whose sputum smear test was negative,and the positive rate were 39.62%(42/106)and 26.42%(28/106)respectively(P<0.05).Conclusion Compared to the traditional L-J solid culture system,BacT/ALERT 3D liquid culture system can shorten detection time and improve the positive detection rate of MTB in specimens with low concentration.

A pilot study on the correlation of tongue manifestation with the site of cerebral infarction in patients with stroke / 中国结合医学杂志

<p><b>OBJECTIVE</b>To discuss the correlation of tongue manifestation with the site of cerebral infarction in patients with acute cerebral infarction.</p><p><b>METHODS</b>From March 2008 to February 2009, 200 cases of hospitalized patients with first unilateral cerebral infarction were chosen in the Department of Neurology, Xuanwu Hospital. The correlation of different tongue color, fur texture, fur color with the site of cerebral infarction was analyzed.</p><p><b>RESULTS</b>The site of cerebral infarction in patients were compared between different tongue color by Chisquare test (P=0.314), and further correspondence analysis demonstrated that there was correlation between red tongue and cortical-subcortical infarction group. The site of cerebral infarction in patients were compared between thick fur group and thin fur group, cortical-subcortical infarction occurred more frequently in the former (P=0.0008). The site of cerebral infarction in patients were compared between dry fur group, moist fur group and smooth fur group, correspondence analysis demonstrated there was correlation between dry fur and cortical-subcortical group. The site of cerebral infarction in the patients were compared between white fur group, white-yellow fur group and yellow fur group (P=0.010), and correspondence analysis demonstrated there was correlation between white fur and brainstem infarction; white-yellow fur has relationship with cortical infarction; subcortical infarction was weakly related with white-yellow fur; there was closer relationship between yellow fur and cortical-subcortical infarction.</p><p><b>CONCLUSION</b>The change of tongue manifestation was associated with the site of cerebral infarction in patients, providing a new combining site for diagnosing cerebrovascular diseases by integrative medicine.</p>

Combined effects of 50 Hz magnetic field and magnetic nanoparticles on the proliferation and apoptosis of PC12 cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the bioeffects of extremely low frequency (ELF) magnetic field (MF) (50 Hz, 400 μT) and magnetic nanoparticles (MNPs) via cytotoxicity and apoptosis assays on PC12 cells.</p><p><b>METHODS</b>MNPs modified by SiO₂ (MNP-SiO₂) were characterized by transmission electron microscopy (TEM), dynamic light scattering and hysteresis loop measurement. PC12 cells were administrated with MNP-SiO2 with or without MF exposure for 48 h. Cytotoxicity and apoptosis were evaluated with MTT assay and annexin V-FITC/PI staining, respectively. The morphology and uptake of MNP-SiO₂ were determined by TEM. MF simulation was performed by Ansoft Maxwell based on the finite element method.</p><p><b>RESULTS</b>MNP-SiO₂ were identified as ~20 nm (diameter) ferromagnetic particles. MNP-SiO₂ reduced cell viability in a dose-dependent manner. MF also reduced cell viability with increasing concentrations of MNP-SiO₂. MNP-SiO₂ alone did not cause apoptosis in PC12 cells; instead, the proportion of apoptotic cells increased significantly under MF exposure and increasing doses of MNP-SiO₂. MNP-SiO₂ could be ingested and then cause a slight change in cell morphology.</p><p><b>CONCLUSION</b>Combined exposure of MF and MNP-SiO₂ resulted in remarkable cytotoxicity and increased apoptosis in PC12 cells. The results suggested that MF exposure could strengthen the MF of MNPs, which may enhance the bioeffects of ELF MF.</p>

Expression of death-associated protein kinase gene and methylation status of promoter region in acute leukemia / 中国实验血液学杂志

This study was purpose to investigate the expression of death-associated protein kinase (DAPK) gene in acute leukemia (AL) patients and the methylation status of its promoter region through experiments of DAPK methylation and expression, and to analyze the relation between them. The expression of DAPK gene in leukemia cells and normal bone marrow cells was detected by RT-PCR; the methylation status of DAPK gene promoter region in cells from AL patients and leukemia cell lines HL-60 and U937 was detected by nested methylation specific PCR (n-MSP); 2 randomly primers selected from randomly amplified products of second round nMS-PCR were cloned and sequenced in professional company. The results showed that the DAPK gene expressed in bone marrow specimens of all 10 normal controls, with average value of expression 0.92 ± 0.18, while the average value of DAPK expression in bone marrow specimens of AL patients was 0.61 ± 0.40 which was lower than that in normal controls (P < 0.05). The low or deletion of DAPK mRNA expression were found in bone marrow specimens of 9/17 (52.94%) cases of ALL and 42/102 (41.18%) cases of AML. The cell line U937 showed normal expression of DAPK gene, while cell line HL-60 showed the expression detection of DAPK gene. The methylation of DAPK promoter region existed in 33 out of bone marrow specimens of 102 AML patients and in 8 out of bone marrow specimens of 17 ALL patients, the methylation rates were 32.4% (33/102) and 47% respectively. The DAPK promoter region in bone marrow of 7 normal controls was unmethylated, while DAPK promoter region in U937 cells and HL-60 cells were unmethylated and methylated respectively. The DAPK mRNA expression in ALL and AML patients significantly negatively correlated with the methylation of its promoter region (r = -0.855, P < 0.05, in AML patients and r = -0.343, P < 0.05, in AML patients) suggesting the close relationship between them. It is concluded that the methylation of DAPK gene promoter region relates with abnormal expression or detection of DAPK mRNA in AL patients.

A pilot study on the relationship between tongue manifestation and the degree of neurological impairment in patients with acute cerebral infarction / 中国结合医学杂志

<p><b>OBJECTIVE</b>To discuss the relationship between tongue manifestation and the degree of neurological impairment in the patients with acute cerebral infarction.</p><p><b>METHODS</b>Two hundred patients with first unilateral cerebral infarction were recruited. The relationship between different tongue manifestation and National Institute of Health Stroke Scale (NIHSS) were analyzed.</p><p><b>RESULTS</b>NIHSS scores in the patients from different tongue color groups were analyzed and further analysis demonstrated that the NIHSS score was higher in the patients with red or bluish-purple tongue than that of those with the pink (P <0.01). On tongue fur, the NIHSS score in the patients with thick fur was higher than that of those with the thin (P=0.003). NIHSS score in patients with slippery, moist or dry fur was significant different (P=0.003), Further analysis demonstrated that the NIHSS score was higher in the patients with dry fur than that of those with moist fur, and had statistical significance (P=0.01). The NIHSS score was higher in patients from greasy fur group than that of the non-greasy (P=0.002). There was significant difference of NHISS score in the patients with different fur color (P=0.000), and further analysis demonstrated that the NHISS score in white-yellow, yellow fur group were higher than that of the white (P=0.06 or 0.000).</p><p><b>CONCLUSION</b>The changes of tongue manifestation might be associated with the degree of neurological impairment in the patients with acute cerebral infarction.</p>

Study on the correlation of tongue manifestation with fibrinogen and neutrophil in acute cerebral infarction patients / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the correlation of tongue manifestation with the fibrinogen level and the neutrophil count in blood of acute cerebral infarction patients.</p><p><b>METHODS</b>A total of 200 patients with first unilateral cerebral infarction in Neurology Department of Xuanwu Hospital from March, 2008 to February, 2009 were recruited in this study. The correlation of the tongue fur color and texture with the blood fibrinogen level and the neutrophil count was analyzed in these patients.</p><p><b>RESULTS</b>The level of fibrinogen and neutrophil count in thick fur group were significantly higher than that in thin fur group (P<0.05). There was no statistical difference in the level of fibrinogen and neutrophil count found between moist fur and dry fur. Statistical significance existed in the level of fibrinogen between the greasy tongue fur group and non-greasy tongue fur group (P<0.05). The level of fibrinogen and the neutrophil count were compared among different fur color groups, revealing that the level of fibrinogen in yellowish fur group was higher than that of white fur group and normal value with statistical significance (P<0.05) with neutrophil count in yellowish fur group being significantly higher than that in white fur group.</p><p><b>CONCLUSION</b>Our results suggested that the change of tongue manifestation was associated with the level of fibrinogen and the neutrophil count in the blood of cerebral infarction patients.</p>

HIF-1 signal pathway in cellular response to hypoxia / 浙江大学学报·医学版

HIF-1 is composed of HIF-1α and HIF-1β subunits. It promotes target genes transcription under hypoxia and plays essential roles in cell development, physiological adaptations, and pathological processes. In the past 10 years, the research on signaling pathways of HIF-1 in response to cell hypoxia stress, especially on HIF-1α-mediated gene transcription has made great progress.

P27(Kip1), cyclin E and endogenous TGF-beta1 changes in apoptosis of NB4 cells induced by As(2)O(3) and/or TGF-beta1 and their significance / 中国实验血液学杂志

This study was aimed to investigate the effects of arsenic trioxide (As(2)O(3)) and/or transforming growth factor-beta1 (TGF-beta1)on cell apoptosis and the changes of P27(Kip1), cyclin E and endogenous TGF-beta1 mRNA levels in NB4 cells. As(2)O(3) cytotoxicity to NB4 cells and the IC(50) were assayed with MTT, the apoptotic morphological changes were observed by Wright-Giemsa staining; the cell cycle and apoptosis were detected with flow cytometry. Semiquantitative RT-PCR was used to examine P27(Kip1), cyclin E and endogenous TGF-beta1 mRNA levels. The results showed that the As(2)O(3) and TGF-beta1 significantly suppressed the growth of NB4 cells, and promoted the apoptosis of these cells. The growth inhibition and apoptosis of NB4 cells treated with As(2)O(3) were in dose-and time-dependent manners. IC(50) were about 12 micromol/L for 24 hours, about 5 micromol/L for 48 hours, and about 3 micromol/L for 72 hours respectively. Cell cycle arrest in NB4 cells was induced by As(2)O(3) and/or TGF-beta1. The arrest of NB4 cells treated by 5 micromol/L As(2)O(3) was in G(2)/M phase, and 5 ng/ml TGF-beta1 in G(1) phase. However, the arrest of NB4 cells caused by combination of As(2)O(3) and TGF-beta1 was in S phase. After treating with As(2)O(3), P27(Kip1) and endogenous TGF-beta1 mRNA expressions of NB4 cells were up-regulated, and cyclin E mRNA expression was down-regulated. When NB4 cells were treated with TGF-beta1 alone, P27(Kip1) and cyclin E mRNA expressions were the same as that treated by As(2)O(3). Exogenous TGF-beta1 enhanced the above effects of As(2)O(3) in combination group. It is concluded that As(2)O(3) and TGF-beta1 are able to induce apoptosis and cell cycle abnormal distribution in NB4 cells. As(2)O(3) and exogenous TGF-beta1 may up-regulate endogenous TGF-beta1, which induce apoptosis of NB4 cells through consequently high expression of P27(Kip1). TGF-beta1 may lead to cell cycle arrest by inhibiting the expression of cyclin E directly, or by the activity of cyclin E through the increased expression of P27(Kip1).

Role of P27(Kip1) and TGF-beta1 in APL cell apoptosis induced by As(2)O(3) / 中国实验血液学杂志

The aim of study was to investigate the effects of arsenic trioxide (As(2)O(3)) on cell cycle and apoptosis of APL cells, as well as changes of P27(Kip1), endogenous TGF-beta1, cyclin E and bcl-2, and to explore the relationship between expression of P27(Kip1) and apoptosis induced by As(2)O(3). The apoptosis and cell cycle changes of APL cells treated with As(2)O(3) were detected by morphology and flow cytometry respectively, the protein and mRNA expressions of P27(Kip1), TGF-beta1, cyclin E and BCL-2 were measured by immunohistochemistry and RT-PCR. The results indicated that As(2)O(3) induced APL cell apoptosis in vitro, and cell cycle was arrested at G(1) phase. Apoptotic cells induced by As(2)O(3) 1, 5 and 10 micromol/L for 24 hours were 1.42%, 4.57% and 10.67% respectively; the proportion of apoptotic cells induced by As(2)O(3) of same concentrations for 48 hours increased to 8.92%, 16.07% and 18.90% respectively; the cells induced by As(2)O(3) for 72 hours were mainly in debris. Protein and mRNA expressions of P27(Kip1) and TGF-beta1 of APL cells after treatment with As(2)O(3) increased, accompanying with decrease of cyclin E, bcl-2 protein and mRNA expressions. Apoptotic cells were related to the expressions of P27(Kip1) (r(mRNA) = 0.55, p < 0.05) and TGF-beta1 (r(mRNA) = 0.51, p < 0.05). There was positive correlation between the expression of TGF-beta1 and of P27(Kip1) (r(mRNA) = 0.31, p < 0.05). It is concluded that the apoptosis of APL cells is induced by As(2)O(3), and the cell cycle is arrested at G(1) phase. The expression of P27(Kip1) is closely related to the extent of apoptosis induced by As(2)O(3). Apoptosis of APL cells induced by As(2)O(3) may be caused by up-regulating TGF-beta1 and P27(Kip1), which is antagonistic to cyclin E and BLC-2, leading to arrest of cell cycle at G(1) phase.

Removing phlegm and dispelling stasis method combined with Western medicine for treatment of cerebrovascular stenosis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of removing phlegm and dispelling stasis method (RPDSM) combined with Western medicine for treatment of cerebrovascular stenosis.</p><p><b>METHODS</b>Seventy enrolled patients were randomly assigned to 3 groups: the Western medicine (WM) group, the integrative medicine (IM) group, and the traditional Chinese medicine (TCM) group. The 21 patients in the WM group were treated with Western medicine as aspirin, Clopidogrel, statins, etc.; the 23 patients in the TCM group were treated with Chinese drugs using the patent preparation Dahuang Zhechong Pill and Tianma Duzhong Capsule as the basic drugs, and supplemented by self-formulated decoctions, selected according to their syndrome types (Tanshi Recipe for dampness-phlegm syndrome, Tanhuo Recipe for fire-phlegm syndrome, Qixu Recipe for qi-deficiency syndrome, and Yang-kang Recipe for yang-excess syndrome); and the 26 patients in the IM group were treated by both TCM and WM with the same drugs and doses mentioned above. The course of treatment was 3 months, and all patients received at least 2 courses in succession. Changes in clinical symptoms and TCM syndrome, levels of C-reactive protein (CRP), platelet aggregation rate (PAR) and fibrinogen (Fib), as well as pictures of medical imaging were observed to evaluate cerebrovascular stenosis.</p><p><b>RESULTS</b>After 6-month treatment, the blood levels of CRP, PAR and Fib were lowered and the number of moderate and severe stenosed vessel lessened in all the three groups (all P < 0.05). The respective total effective rate in the WM, TCM and IM group was 42.9% (9/21 cases), 39.1% (9/23 cases) and 61.5% (16/26 cases), no significant difference was shown among them.</p><p><b>CONCLUSION</b>The integrative Chinese traditional and Western medical treatment for cerebrovascular stenosis shows an increasing trend in improving clinical efficacy and laboratory indexes, combared with pure Western or Chinese medical treatment.</p>

Effect of bufalin-inducing apoptosis on Bcl-2 and PKC in HL-60 cells / 中国实验血液学杂志

Previous study revealed that bufalin can inhibit proliferation, and induce apoptosis in some human cancer cell lines. However, the mechanism of its anticancer effect has not been fully understood. The present study was designed to investigate the effects of bufalin-induced apoptosis on Bcl-2 and PKC in human leukemic HL-60 cells. The cell viability was determined by trypan blue dye exclusion. The apoptosis was detected by morphology, flow cytometry and DNA agarose gel electrophoresis. The expressions of Bcl-2 and PKC were analyzed by Western blot, and activity of PKC was assayed by [gamma-(32)P] isotope incorporation method. The results showed as follows: (1) proliferation of HL-60 cells was inhibited by bufalin and the IC(50) at 24, 48, 72 hours were (25.8 +/- 2.1), (8.0 +/- 1.2) and (2.3 +/- 0.3) nmol/L, respectively. (2) apoptosis of HL-60 cells was induced when the cells were treated with bufalin at concentration of 50 nmol/L for 24 hours. (3) compared with control, treatment with bufalin at concentration of 50 nmol/L for 6 - 24 hours resulted in downregulation of protein expression, decrease of phosphorylation, and cleavage of Bcl-2, simultaneously. (4) the activity of total PKC was unchanged when HL-60 cells were exposed to 1 - 100 nmol/L bufalin for 30 minutes, but PKCbetaII underwent translocation from cytosol to membrane. It is concluded that apoptosis induced by bufalin is associated with downregulation of protein expression, dephosphorylation, and cleavage of Bcl-2 in HL-60 cells.

Mutation analysis in families with 21-hydroxylase deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the steroid 21-hydroxylase gene (CYP21) mutations in families with 21-hydroxylase deficiency (21-OHD).</p><p><b>METHODS</b>The CYP21 gene mutations were detected in four patients with 21-hydroxylase deficiency and their relatives. The genomic DNA of the patients was isolated from whole blood.Two pairs of primers were used to amplify the CYP21 gene. The amplified PCR products were purified by agarose gel and then directly sequenced.</p><p><b>RESULTS</b>Six kinds of mutations were found. In the first family, the patient was a compound heterozygote carrying four different mutations (cluster E6, Q318X, A391T, P459H) onCYP21 gene, three mutations (cluster E6, Q318X, A391T) were on her maternal allele, a novel mutation was found:P459H. It located at codon 459 in exon 10 and changing a proline (CCC) to a histidine (CAC), and A391T was a rare mutation. In the second family, two kinds of mutations were found:cluster E6 and R483W. R483W was also a rare mutation. In the third family, the sequencing of the CYP21 gene of two patients revealed a homozygous T to A transition in codon 172 leading to substitution of isoleucine by asparagine (I172N).</p><p><b>CONCLUSION</b>Six kinds of mutations were found in three families with 21-hydroxylase deficiency. Using DNA sequencing we have identified a novel mutation (P459H) and two rare mutations (A391T, R483W) of the CYP21 gene. Although microconversion events are the main cause of mutations in the CYP21 gene, random mutations can also be the cause of 21-hydroxylase deficiency.</p>