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.

Establishment of induced pluripotent stem cell model of Aicardi-Goutières Syndrome mutated in TREX1 / 中华预防医学杂志

To establish and identify induced pluripotent stem cells (iPSCs) derived from patients with Aicardi-Goutières syndrome (AGS) with TREX1 gene 667G>A mutation, and obtain a specific induced pluripotent stem cell model for Aicardi-Goutières syndrome (AGS-iPSCs). A 3-year-old male child with Aicardi-Goutieres syndrome was admitted to Zhongshan People's Hospital in December 2020. After obtaining the informed consent of the patient's family members, 5 ml peripheral blood samples from the patient were collected, and mononuclear cells were isolated. Then,the peripheral blood mononuclear cells(PBMCs) were transduced with OCT3/4, SOX2, c-Myc and Klf4 by using Sendai virus, and PBMCs were reprogrammed into iPSCs. The pluripotency and differentiation ability of the cells were identified by cellular morphological analysis, real-time PCR, alkaline phosphatase staining (AP), immunofluorescence, teratoma formation experiments in mice. The results showed that the induced pluripotent stem cell line of Aicardi-Goutieres syndrome was successfully constructed and showed typical embryonic stem-like morphology after stable passage, RT-PCR showed mRNA expression of stem cell markers, AP staining was positive, OCT4, SOX2, NANOG, SSEA4, TRA-1-81 and TRA-1-60 pluripotency marker proteins were strongly expressed. In vivo teratoma formation experiments showed that iPSCs differentiate into the ectoderm (neural tube like tissue), mesoderm (vascular wall tissue) and endoderm (glandular tissue). Karyotype analysis also confirmed that iPSCs still maintained the original karyotype (46, XY). In conclusion, induced pluripotent stem cell line for Aicardi-Goutières syndrome was successfully established using Sendai virus, which provided an important model platform for studying the pathogenesis of the disease and for drug screening.

Nimotuzumab plus induction chemotherapy followed by radiotherapy/concurrent chemoradiotherapy plus nimotuzumab for locally advanced nasopharyngeal carcinoma: protocol of a multicentre, open-label, single-arm, prospective phase II trial.

Epidermal growth factor receptor (EGFR) is a therapeutic target in nasopharyngeal carcinoma (NPC). The optimal combined modality of optimal combined modality of anti--EGFR monoclonal antibodies, induction chemotherapy (ICT), concurrent chemotherapy and radiotherapy for NPC remains poorly defined. None of previous studies have developed subsequent treatment strategies on the basis of stratification according to the efficacy following ICT plus anti-EGFR mAbs. This study aims to increase treatment intensity for patients with poor efficacy of ICT and reduce treatment toxicity for patients with favourable efficacy of ICT by assessing whether the efficacy of this treatment regimen is non-inferior to ICT plus concurrent chemoradiotherapy (historic controls). INTRODUCTION: METHODS AND ANALYSIS: Pathology-confirmed WHO type II/III NPC patients at clinical stage III-IVA (eighth American Joint Committee on Cancer/Union for International Cancer Control staging system) will be included in the study. They will receive ICT plus nimotuzumab (NTZ), followed by radiotherapy plus NTZ or concurrent chemoradiotherapy plus NTZ (stratified based on the efficacy of ICT plus NTZ). The primary endpoint is 3-year failure-free survival rate; while the secondary endpoints are 3-year overall survival rate, distant metastasis-free survival rate and locoregional recurrence-free survival rate, and short-term remission rate of tumour and treatment toxicity. ETHICS AND DISSEMINATION: The study protocol has been approved by the Ethics Committee of the Second Affiliated Hospital of Nanchang University. Our findings will be disseminated in a peer-reviewed journal. Implementation strategies are in place to ensure privacy and confidentiality of participants. TRIAL REGISTRATION NUMBER: ChiCTR2000041139.

Evaluation of Burnout Among Anesthesiologists Working in Tibet, China: Altitude and Attitude / 中国医学科学杂志(英文版)

Objective Burnout is a triad of emotional exhaustion, depersonalization, and reduced personal accomplishment resulting from job stress. Although with distinct regional and cultural characteristics, burnout among anesthesiologists in the Tibet has not been described. This study aimed to explore the prevalence of burnout among anesthesiologists in Tibet and its associated factors. Methods A cross-sectional survey was conducted in Tibet, China, with an anonymous questionnaire. Social-demographic characteristics, work status, three dimensions of burnout assessed by the Maslach Burnout Inventory-Human Service Survey were collected and analyzed. Results A total of 133 individuals from 17 hospitals completed the survey from March to June 2018. The prevalence of moderate- to high-level of emotional exhaustion, depersonalization, and burnout in personal accomplishment was 65.4% (95%

Assessing Empathy in Anesthesia Residents with the Chinese Version of the Consultation and Relational Empathy: A Pilot Standardized Patient Program / 中国医学科学杂志(英文版)

Objectives To validate the reliability of the Chinese version of the Consultation and Relational Empathy (CARE) in physician-standardized patient (SP) encounter. We also tried to examine the agreement between video-based ratings and in-room ratings, as well as the agreement between the faculty ratings and SP ratings. Methods The CARE was translated into Chinese. Forty-eight anesthesia residents were recruited to make preoperative interview in SP-counter. Performance of each resident was graded by in-room raters, video raters and SP raters. Consistency between different raters was examined. Results The Chinese-CARE measure demonstrated high scale reliability with a Cronbach's alpha value of 0.95 and high consistency in the in-room ratings in intraclass correlation (coefficient=0.888,

Characteristics and Management of High-risk Maternal Patients in General Tertiary Hospital / 中国医学科学院学报

Objective To analyze the characteristics of high-risk maternal patients and evaluate the multidisciplinary medical care system we established correspondingly. Method We collected and analyzed the medical records of high-risk maternal patients who received medical care from January 1,2017 to December 31,2020 in Peking Union Medical College Hospital. Results Ninety-eight high-risk maternal patients were included in this study,and 84.7%(83/98)of them were combined with different severe systemic diseases.Under the multidisciplinary medical care system,91 patients showed improved conditions and were discharged,and the other 7 cases had poor prognosis. Conclusions General tertiary hospitals in Beijing are receiving maternal patients with more high-risk complications.Considering the high risk and diverse diseases of maternal patients admitted to our hospital,we established a medical care system composed of a multidisciplinary panel of experts for high-risk maternal patients to improve the medical care and prognosis of the patients with high efficiency.

Efficacy of Oxycodone Hydrochloride for Patient-controlled Intravenous Analgesia after General Surgery / 中国医学科学院学报

To evaluate the value of oxycodone hydrochloride for postoperative pain management in patients undergoing patient-controlled intravenous analgesia(PCIA). The medical records on postoperative pain management in our department from January 1 to June 30,2018,were retrospectively analyzed.Totally 136 patients were assigned into oxycodone,sufentanil,or morphine groups according to the opioid used in the PCIA.Patients were assessed for postoperative pain severity(scored with NRS)and adverse reactions 24,36,and 48 hours after surgery.The area under curve(AUC)was calculated. The score of pain at exercise was significantly lower in the oxycodone group(2.2±2.4)than in the sufentanil group(3.4±2.1)(=0.305,=0.0126)or the morphine group(3.4±1.7)(=0.104,=0.0277)36 hours after surgery.AUC at rest was significantly lower in the oxycodone and morphine groups than in the sufentanil group(29.00,27.00,and 40.01,respectively);in contrast,AUC at exercise was significantly lower in the oxycodone group(63.17)than in the sufentanil and morphine groups(82.00 and 80.93,respectively).The consumption of opioids was significantly higher in the sufentanil group[(37.2±16.1),(46.1±24.3),(64.4±33.4)mg]than in the oxycodone group[(20.4±14.8)(=3.571,=0.001),(24.2±16.1)(=4.63,<0.0001),(34.4±25.1)mg(=6.409,<0.0001)]or the morphine group[(16.6±11.7)(=4.233,<0.0001),(20.5±14.1)(=5.250,<0.0001),(28.8±19.0)mg(=7.354,<0.0001)]24,36,48 hours after surgery.The oxycodone group experienced less vomiting(=11.360,=0.003)and early termination of PCIA(=7.914,=0.019)compared with the other two groups. Oxycodone can be used for postoperative PCIA.It can alleviate a variety of postoperative pain,with superior analgesic efficiency and safety to sufentanil and morphine.

The high pCR rate of sandwich neoadjuvant treatment in locally advanced rectal cancer may translate into a better long-term survival benefit: 5-year outcome of a Phase II clinical trial.

BACKGROUND: In a Phase II clinical trial, we reported the effectiveness and safety of a sandwich neoadjuvant treatment based on a modified oxaliplatin plus capecitabine (XELOX) regimen for locally advanced rectal cancer (LARC). The pathologic complete response (pCR) rate was 42.2%, and no patient presented Grade 4 acute toxicities. This study was performed to evaluate whether the high pCR rate could translate into an improved long-term survival benefit by analyzing the 5-year follow-up results of the trial. METHODS: Fifty-one patients with LARC were initially enrolled in the trial. Of these, 2 cases were eliminated due to distant metastasis before treatment. In addition, 4 cases were eliminated for refusing surgery after neoadjuvant chemoradiotherapy (NACRT). Finally, a total of 45 patients were treated with the sandwich NACRT plus total mesorectal excision. We followed up these patients and calculated their overall survival (OS) and disease-free survival (DFS) through a Kaplan-Meier approach. A log-rank test and multivariate survival analysis based on a Cox proportional hazard model were performed to explore the risk factors influencing distant metastasis. RESULTS: The median follow-up time was 60.8 months, and among the 45 patients analyzed, 1 (2.2%) patient suffered local recurrence, and 9 (20.0%) suffered distant metastasis. The 3-year OS and DFS were 95.6% and 84.4%, respectively. In addition, the 5-year OS and DFS were 91.1% and 80.0%, respectively. In the multivariate analysis, postsurgical pathological N stage and carbohydrate antigen 19-9 before treatment maintained statistical significance on distant metastasis. CONCLUSIONS: The sandwich NACRT with XELOX regimen might reduce distant metastasis and improve the survival of LARC patients. However, long-term benefits should be verified through further Phase III clinical trials.

Apolipoprotein A-I Is a Prognosticator of Nasopharyngeal Carcinoma in the Era of Intensity-modulated Radiotherapy.

Background: In the era of intensity-modulated radiotherapy (IMRT), distant metastasis remains the major cause of death from nasopharyngeal carcinoma (NPC). This study aimed to evaluate the clinical value of pretreatment serum lipid profiles in predicting clinical outcome of NPC. Methodology / Principal Findings: A total of 1927 consecutive patients who had untreated NPC and completed radical IMRT between Jan. 2010 and Dec. 2011 were retrospectively reviewed. Pretreatment serum lipid indexes including total cholesterol, triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, apolipoprotein A-I (apoAI) and apolipoprotein B were analyzed for their association with survivals, together with the clinical features (age, sex, pathological type, anemia, chemotherapy sequence and Epstein-Barr virus deoxyribonucleic acid). Hazard ratio (HR) and 95% confidence interval (CI) were calculated for each independent prognosticator. After univariate and multivariate survival analysis, low apoAI level (< 1.125 mmol/L) appeared to predict poor 5-year overall survival (OS), disease-free survival (DFS) and distant-metastasis-free survival (DMFS).The HRs were 1.549 (95% CI, 1.137-2.109), 1.293 (95% CI, 1.047-1.597) and 1.288 (95% CI, 1.022-1.623), respectively. Subgroup survival analysis showed that the apoAI maintained predicting independence for OS, DFS and DMFS in patients with locally advanced NPC, even in those treated with concurrent chemoradiotherapy. Conclusions / Significance: NPC patient with low serum level of pretreatment apoAI might be at risk of distant metastasis. Treatment aiming to eradicate distant metastasis might improve survival of these patients.

CCR6 Is a Predicting Biomarker of Radiosensitivity and Potential Target of Radiosensitization in Rectal Cancer.

PURPOSE: This study aimed to explore the functions and mechanisms of C-C motif chemokine receptor 6 (CCR6), a gene associated with progression and metastasis of colorectal cancer (CRC), in radiosensitivity of rectal cancer (RC). MATERIALS AND METHODS: RNA sequencing and immunohistochemical analysis on CCR6 expression were performed in pretreatment tissues of RC patients exhibiting different therapeutic effects of radiotherapy. Colonogenic survival assay was conducted in different CRC cell lines to assess their radiosensitivity. And the impact of CCR6 expression on radiosensitivity was validated through RNA interference. The DNA damage repair (DDR) abilities of cell lines with different CCR6 expression were evaluated through immunofluorescence-based γH2AX quantification. RESULTS: The CCR6 mRNA level was higher in patients without pathologic complete remission (pCR) than in those with pCR (fold changed, 2.11; p=0.004). High-level expression of CCR6 protein was more common in the bad responders than in the good responders (76.3% vs. 37.5%, p < 0.001). The CRC cell lines with higher CCR6 expression (LoVo and sw480) appeared to be more radioresistant, compared with the sw620 cell line which had lower CCR6 expression. CCR6 knockdown made the LoVo cells more sensitive to ionizing radiation (sensitization enhancement ratio, 1.738; p < 0.001), and decreased their DDR efficiency. CONCLUSION: CCR6 might affect the RC radiosensitivity through DDR process. These findings supported CCR6 as a predicting biomarker of radiosensitivity and a potential target of radiosensitization for RC patients.

Application of standardized patients in the teaching of preoperative evaluations of anesthesia residents / 基础医学与临床

Standardized patients (SPs) are trained to portray a specific patient in a consistent, standardized fash-ion. SPs are widely used in many medical schools,and its efficacy for education and evaluation in the medical com-munity has been well established. Preoperative evaluation by anesthesiologist is a critical aspect of patient safety. But there are several defects in the current anesthesia education of preoperative evaluation and management. The residents are not provided with enough skills that are needed in the clinical practice and this sbortage may be com-pansated by SPs simulation. As a supplement of traditional teaching methods,SPs may improve the efficiency in the teaching of preoperative evaluation and enhance the competence of anesthesia residents.

Factors Influencing Regional Cerebral Oxygen Saturation during One-lung Ventilation in Thoracic Surgery / 中国医学科学院学报

Objective To identify factors influencing regional cerebral oxygen saturation (rScO) during one-lung ventilation (OLV) in thoracic surgery. Methods Totally 33 patients with an ASA physical status of 1-3 scheduled for elective thoracic surgery with one-lung ventilation under general anesthesia were recruited. After anesthesia was induced with propofol,fentanyl/sufentanil,and rocuronium. All patients received balanced anesthesia using sevoflurane. During OLV,volume-controlled ventilation was used with a tidal volume of 6-7 ml/kg and an inspiration:expiration ratio of 1:1.5. The ventilator frequency was adjusted with a target end-tidal carbon dioxide partial pressure (PetCO) between 35 mmHg and 45 mmHg. During the anesthesia,patients were maintained at a pulse oxygen saturation (SpO) of>90%,systolic blood pressure (SBP) of>90 mmHg (or reducing no more than 30% of the basic values),heart rate (HR) of>50 beat/min,and hemoglobin concentration of>90 g/L. Changes of rScOwere monitored with FORESIGHT probes by specialized researchers. Patients were classified into low rScO(L-rScO) group (n=10) or high rScO(H-rScO)group(n=23) according to whether the lowest intraoperative rScOwas under 65% or 15% lower than the baseline values. We compared gender,age,body mass index (BMI),intraoperative hemoglobin level,and the values of peak airway pressure (Ppeak),SBP,PetCO,and SpOwhen rScOdropped to the lowest level between these two groups. Results Statistically higher Ppeak and lower SBP were noted in the L-rScOgroup compared with H-rScOgroup (P=0.028,P=0.046). SpOwas lower in the L-rScOgroup compared with H-rScOgroup,but the difference was not statistically significant (P=0.421). There was also no significant difference between the two groups according to age,BMI,SpO,PetCO,or hemoglobin level. Ppeak appeared to be a risk factor for rScOreduction during OLV,as shown by unconditioned Logistic regression analysis. Conclusion During OLV in thoracic surgery,Ppeak is a risk factor for rScOreduction.

Effect of Inhibiting and Activating Wnt Signalling Pathway on NSC67657-inducing Monocytic Differentiation of HL-60 Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect of inhibiting and activating Wnt signalling pathway on monocyte differentiation of HL-60 cells induced with a new steroidal drug NSC67657 and its possible mechamism.</p><p><b>METHODS</b>The HL-60 cells were treated with 5, 10 and 20 µmol/L XAV-939 (inhibitor of Wnt signalling pathway) for 3 days, and with 10, 20 and 30 mmol/L LiCl (activator of Wnt signalling pathway) for 1 day; the expression levels of down-stream genes and proteins of Wnt signolling pathway were detected by RT-PCR and Western blot, respectively; the expression of cell surface differentiation antigen CD14 and early apoptosis of HL-60 cells was detected by flow cytometry, moreover the most suitable concentration of Wnt inhibitor and activator for HL-60 cells was determined. Then the HL-60 cells with inhibited and activated Wnt pathway were treated with NSC67657 of 10 µmol/L for 3 days; the expression levels of CD14 and down-stream target proteins of Wnt signalling pathway in blank control (culture mediam) group, simple NSC67657-treated group, NSC67657 combined with inhibitor group and NSC67657 combined activator group were compared and analyzed.</p><p><b>RESULTS</b>20 µmol/L XAV-939 and 20 mmol/L LiCl could effectively inhibit and activate Wnt signalling pathway of HL-60 cells respectively, could significantly down- and up-regulate the expression of cyclinD1, TCF1 and c-Jun genes (P < 0.05) and proteins (P < 0.05); moreover, the number of CD10(+) HL-60 cells in these conditions was below 1%, no early apoptosis of HL-60 cells was found. In the simple NSC67657-treated groups, the expression of cyclinD1, TCF1 and c-Jun proteins was down-regulated (P < 0.05), and the percentage of CD14(+) HL-60 cells accounted for 62.13 ± 9.44; after the HL-60 cells were treated with XAV-939, the NSC67657 could more significantly down-regulate the expression of cyclinD1, TCF1 and c-Jun proteins and the percentage of CD14(+) HL-60 cell accounted for 84.17 ± 5.39%, as compared with simple NSC67657-treated group; as compared with blank controls group, the expression of cyclinD1, TCF1 and c-Jun proteins was more obviously down-regulated and the percentage of CD14(+) HL-60 cells decreased to 33.99 ± 8.37% in NSC67657 combined LiC1 streated group, but which were higher than those in simple NSC67657-treated group (P < 0.05).</p><p><b>CONCLUSION</b>20 µmol/L XAV-939 and 20 mmol/L LiCl as effective inhabitor and activator of Wnt signalling pathway respectively can significantly down- and up-regulate the expression of Wnt down-stream pathway target genes and proteins. The influence of XAV-939 and LiC1 on differentiation of HL-60 cells induced by NSC67657 suggests that Wnt signalling pathway plays a key role in monocyte differentiction of HL-60 cells induced by NSC67657.</p>

Expressing trend of NME3 protein in acute myeloid leukemia HL-60 cells and patients' bone marrow / 中国实验血液学杂志

To verify the differential expression of non-metastasis cell 3 (NME3) protein in HL-60 cells when they were induced to differentiate into monocyte and granulocyte like cells, and study its value in diagnosis of acute myeloid leukemia, all-trans retinoic acid (ATRA) and a new steroidal drug NSC67657 were employed to induce acute myeloid leukemia HL-60 cells into monocyte and granulocyte like cells. Then the cell differentiating direction was observed by chemical staining, the degree of differentiation was determined by surface antigen CD11b/CD14 detection, and the apoptosis was excluded by phosphatidylserine valgus analysis, by which cellular differentiating model was constructed. Furthermore, RT-PCR and Western blot were employed to verify the differentially expression of NME3 before and after differentiation of HL-60 cells. At last, samples from bone marrow nucleated cells of 26 patients with myeloid leukemia, which were diagnosed definitely by clinical doctors, and 5 normal people were chosen. Then the expressing trend of NME3 protein in these testing groups was analyzed by means of comparison. The results showed that ATRA (2 µmol/L for 5 d) and NSC67657 (10 µmol/L for 5 d) could induce HL-60 cells to differentiate into monocyte and granulocyte like cells above 90% without cell apoptosis. The expression of NME3 gene and protein were down-regulated by the inducers, which was accorded with the screening results that was got using proteomics technology in the former research. The expression of NME3 protein in bone marrow from acute myeloid leukemia patients was elevated significantly as compared to normal persons. It is concluded that the expression level of NME3 protein is down-regulated after cellular differentiation, according with the changing trend in leukemia patients, which imply that NME3 protein may be a potential biomarker for diagnosis of acute myeloid leukemia.

Amino-terminal pro-brain natriuretic peptide and brain natriuretic peptide measurements under various detection conditions in patients with chronic heart failure / 中华心血管病杂志

<p><b>OBJECTIVE</b>To find the potential interference factors for the detection of NT-proBNP and BNP in patients with chronic heart failure.</p><p><b>METHODS</b>EP15-A2 issued by Clinical and Laboratory Standards Institute (CLSI) was employed to compare the precision and accuracy of commercial NT-proBNP and BNP analyzer electrochemiluminescence immunoassay system Cobas E601 and chemiluminescence system ADVIA Centaur. Moreover, NT-proBNP and BNP were detected in different time interval and in different interfered sampling conditions (haematolysis, choloplania, lipemia). NT-proBNP and BNP of 203 patients with heart failure or heart failure complicated with acute cerebral infarction were analyzed to find the deviation caused by patients' endogenous factors.</p><p><b>RESULTS</b>The precision and accuracy were comparable for NT-proBNP and BNP detection using Cobas E601 and ADVIA Centaur (total-CV below 2.9% and 3.5%, the deviation from definite value below 2.38% and 3.91%). The most suitable sample type for NT-proBNP and BNP detection was serum and EDTA-anticoagulant plasma. The detection results of NT-proBNP and BNP were comparable for at least 120 min post sampling and not affected by Hb (2 g/L), DB (428 µmol/L) and chyle (2000 FIU). NT-proBNP was significantly higher in heart failure patients complicated with cerebral infarction (P = 0.003) than in heart failure patients. BNP was significantly higher in heart failure grade III patients complicated with cerebral infarction (P < 0.01).</p><p><b>CONCLUSIONS</b>Cobas E601 and ADVIA Centaur supplied satisfactory detection of NT-proBNP and BNP in patients with chronic heart failure with strong anti-interference capacity. The diagnostic value of NT-proBNP and BNP for chronic heart failure should be analyzed objectively in the presence of complicating diseases.</p>

Study on the mechanism of THP-1 cell differentiation imduced by a new steroidal drug NSC67657 / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the potential mechanism of the new steroidal drug NSC67657 induced leukemic cells differentiation.</p><p><b>METHODS</b>Cell proliferation was assayed by MTT assay. Surface antigen CD14 on THP-1 cells treated by NSC67657 at different time different concentration, was detected by flow cytometry (FCM). The expression of beta-catenin- interacting protein 1 (ICAT) gene and protein were detected by RT-PCR and Western blot. Eukaryotic expressing vector pDsRed-ICAT was constructed and transfected into HL60 cell line. FCM, Wright's staining and electronmicroscope were employed to analyse the differentiation of transfected THP-1 cells after they were treated with NSC67657 for 24 hours.</p><p><b>RESULTS</b>The proliferation of THP-1 cells was significantly inhibited by NSC67657 treatment. The level of CD14 expression was elevated in line with the increasing drug concentration and treatment time. 10 µmol/L NSC67657 treatment for five days was the optimal condition for the induction of THP-1 cells differentiation, when the CD14(+) THP-1 cells were more than 90%. Morphological study indentified the THP-1 cells of monocytic differentiation. The eukaryotic expressing vector pDSRed-ICAT was successfully constructed, and almost 90% positive clone could be obtained after G418 screening. Electro-transfection was employed for transfecting the vector into THP-1 cells. After the transfection the expression of ICAT gene and protein was increased. On the NSC67657 treatment, there was not significant difference in CD14 expression on transfected THP-1 cells compared to that on the control groups. After 24 h treatment, the transfected THP-1 cells remained in early differentiated stage.</p><p><b>CONCLUSION</b>NSC67657 can induce THP-1 cell to monocytic differentiation and activate the expression of ICAT gene, but overexpression of ICAT itself is not sufficient to induce such differentiation.</p>

Effect of C/EBPalpha on the monocytic differentiation of HL60 cells induced by NSC67657 / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To figure out the function of C/EBPalpha in the monocytic differentiation of HL60 cells induced by a new steroidal drug NSC67657.</p><p><b>METHODS</b>The differentiation of HL60 cells was induced by NSC67657, and the cell surface antigen CD14 expression was detected by flow cytometry. The gene and protein expressions of CCAAT enhancer binding protein alpha (C/EBPalpha) before and after the induction of cell differentiation were determined by RT-PCR and Western blot. Eukaryotic expressing vector pDsRed-ICAT was constructed and transfected into HL60 cells, and its expression was verified. The effect of C/EBPalpha overexpression in HL60 cells was assessed by MTT assay, Wright's staining and flow cytometry before and after NSC67657 transfection.</p><p><b>RESULTS</b>HL60 cells could be induced into monocytes by 10 micromol/L ATRA within 5 days, and the coverage of CD14 positive cells reached 93.9% after 5 days of drug treatment. The eukaryotic expressing vector was successfully constructed, and over 90% positive clones were obtained after screening by G418 and electrotransfection. The results of proliferative analysis, chemical staining, ultrastructural observation, and CD11b detection confirmed that HL60 cells could be induced into granulocytic differentiation by overexpression of C/EBPalpha protein. Moreover, in the drug treatment group, transfected cells could not be induced into monocytic differentiation, and their granulocytic differentiation was also inhibited.</p><p><b>CONCLUSION</b>The monocytic differentiation of HL60 cells induced by NSC67657 may not be via the regulation by C/EBPalpha protein-mediated signal transduction. However, the overexpression of CEBPalpha may inhibit the process of NSC67657-induced monocytic differentiation in HL60 cells.</p>

Recent advances of Drynaria fortunei / 中国骨伤

Drynaria fortunei (Kunze) is a common medicine in department of orthopaedics and traumaology, more researches about it recently, including basic theory, pharmaco and clinical study.

Effects of matrine on cgi-100 gene expression and proliferation in K562 cells / 中国实验血液学杂志

This study was aimed to explore the expression level of the unknown cgi-100 gene in human leukemia K562 cells treated with matrine, and to investigate effect of cgi-100 on proliferation of K562 cells. The expression level of cgi-100 was detected by RT-PCR in K562 cells before and after being treated with matrine; pIRES2-EGFP/cgi-100 eukaryotic expression vector was constructed by DNA recombinant technique and was introduced into K562 cells by liposome-mediated DNA transfection. The cgi-100 gene expression level, growth-curve, and cell cycle of the modified K562-cgi-100 cells were detected by RT-PCR, Trypan blue staining and FCM. Morphological changes were observed under the optical and electron microscopes. The results indicated that the expression level of cgi-100 decreased in K562 cells treated with matrine. Heterochromatin decreased, euchromatin and the proportion of S phase in K562-cgi-100 cells increased, and cell proliferation enhanced. It is concluded that the expression of cgi-100 mRNA decreased in a dose- and time-dependent manner in the K562 cells treated with matrine and over-expression of cgi-100 elevates the proliferation and the immaturity level of K562-cgi-100 cells.

The expression of IER3IP1 gene in K562 cells treated by matrine and its effect on the cell growth / 中华血液学杂志

<p><b>OBJECTIVES</b>To investigate the expression status of IER3IP1 gene during matrine induced K562 cell differentiation, and to figure out the function of IER3IP1 gene in K562 cell line.</p><p><b>METHODS</b>Trypan-blue staining was used to analyze the growth inhibitory effect of matrine on K562 cells. Semi-quantitative RT-PCR was employed to investigate the expression status of IER3IP1 gene treated with different time and dosage of matrine. The alteration of cellular morphology, cellular proliferation and ultra-microstructure were observed and the cell cycle was detected on the recombinant IER3IP1 gene eukaryotic expression vector eYFP-IER3IP1 plasmid transfected K562 cells (K562/eYFP-IER3IPl).</p><p><b>RESULTS</b>Matrine inhibited the growth of K562 cells and reduced the expression of IER3IP1 gene. The expression level of IER3IP1 gene was transiently increased to three-to-four times in a dose-dependent manner after treated with matrine for 2 - 3 hours. Then, in 6-48 hours it maintained at a low level as compared with the control group. The proliferation rate of the K562/eYFP-IER3IP1 cells significantly slowed down with more cells blocked in G0-G1 phase (P < 0.05). The number of erythroid blast cells began to increase after 24 hours of matrine treatment. At the same time, differentiated erythroid cells could be observed.</p><p><b>CONCLUSIONS</b>Matrine can inhibit the growth of K562 cells, and transiently increase the expression level of IER3IP1 gene in a dose-dependent manner. The sensitivity of K562 cells to matrine maybe increased after being transfected by the eYFP-IER3IP1 plasmid, indicating a possible involvement of the IER3IP1 gene in the early response of the cells to matrine and its possible role in the erythroid cell differentiation.</p>