Preclinical study of LMP1-RNAi-based anti-tumor therapy in EBV-positive nasopharyngeal carcinoma

RNA interference (RNAi) treatment has been proven to be an important therapeutic approach in cancer based on downregulation of target-oncogenes, but its clinical efficacy still needs further investigation. LMP1 is usually presented by Epstein-Barr virus (EBV)-positive tumor cells like EBV-associated nasopharyngeal carcinoma (NPC) and acts as an oncogene in tumorigenesis. However, the mechanism of LMP1 as a proto-oncogene in nasopharyngeal carcinoma is still unclear. Two sequence-specific shRNAs 1 and 2 were designed to target the different nucleotide loci of EBV latent antigen LMP1 gene and a series of in vivo and in vitro experiments were performed to investigate the therapeutic effect of sequence-specific shRNAs targeting LMP1 and its related molecular mechanisms in EBV-positive NPC. LMP1-shRNA2 generated a truncated LMP1 mRNA and protein, whereas LMP1-shRNA1 completely blocked LMP1 mRNA and protein expression. Both LMP1-shRNAs inhibited the proliferation and migration of NPC cells overexpressing LMP1 (NPC-LMP1) as well as the NPC-associated myeloid-derived suppressor cell (MDSC) expansion in vitro. However, LMP1-shRNA2 maintained the immunogenicity of NPC-LMP1 cells, which provoked MHC-class I-dependent T cell recognition. LMP1-shRNAs inhibited tumor growth in nude mice but did not reach statistical significance compared to control groups, while the LDH nanoparticle loaded LMP1-shRNAs and the antigen-specific T cells induced by NPC-LMP1 cells treated with LMP1-shRNA2 significantly reduced tumor growth in vivo. LMP1-RNAi-based anti-tumor therapy could be a new hope for the clinical efficacy of RNAi treatment of tumors like NPC.

New advances in the study of subthreshold micropulse laser photocoagulation for central serous chorioretinopathy / 中华眼底病杂志

Central serous chorioretinopathy (CSC) is a common macular disease, which can lead to the loss of central vision in young patients. Although the disease is self-limited to some extent, there is no shortage of patients with prolonged course and recurrent attacks, eventually leading to irreversible visual impairment. Therefore, taking reasonable treatment in a certain period is particularly important for the visual prognosis of patients. Although thermal laser photocoagulation of leakage points and photodynamic therapy (PDT) have achieved good effects, there are risks of retinal damage and retinal pigment epithelium atrophy. Subthreshold micropulse laser (SMLP) is a kind of subthreshold short pulse laser, which does not cause visible damage to the retina and is safer. In the era of lack of PDT drugs, SMLP has gradually become an important means of clinical treatment for CSC, especially for patients with no obvious leakage point or subfoveal leakage point. An in-depth understanding of the mechanism of action of SMLP and its efficacy and safety in the treatment of CSC is helpful for the promotion and application of SMLP in the clinical treatment of CSC.

Identification of key immune related genes in diabetes retinopathy based on weighted gene co-expression network / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the key genes related to immunity and immune cell infiltration levels in diabetes retinopathy(DR)using bioinformatics.METHODS: Differential expression genes(DEGs)were obtained by “limma” R from Gene Expression Omnibus(GEO)data from September to October 2022, Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)were analyzed, and the infiltration of immune cell types in each sample was calculated based on CIBERSORT algorithm. Weighted gene co-expression network analysis(WGCNA)was used to screen for DEGs in immune-related gene modules. The protein-protein interaction(PPI)network was established by STRING online database and Cytoscape, and the hub genes were screened by MCODE and cytoHubba plug-ins.RESULTS: The results showed that 1 426 up-regulated and 206 down-regulated differential genes were screened, where 7 immune cell types, including B cell naive, Plasma cells, CD4+T cells, T cells regulatory(Tregs), Macrophages M0, Macrophages M1 and Neutrophils were significantly overexpressed(P<0.05), while others were low expressed(P<0.05). After WGCNA, a total of 820 DEGs were found in the modules most related to immunity. After constructing the PPI network, 10 key genes were screened using plug-ins, and two key genes were further screened using the expression amount of each differential gene in PPI: DLGAP5 and AURKB.CONCLUSION: This study used bioinformatics to screen the infiltration of immune cells and key genes related to immunity in patients with DR. These findings may provide evidences for future research, diagnosis, and treatment of DR.

Effect of repeated intravitreal Ranibizumab and Aflibercept injections on the corneal nerves in patients with macular edema / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect of repeated intravitreal injection of ranibizumab and aflibercept on corneal nerve of patients with macular edema.METHODS: A total of 64 patients(64 eyes)enrolled in our hospital from June 2021 to June 2022 were treated with intravitreal injection of anti-vascular endothelial growth factor(VEGF). There were 20 cases(20 eyes)of diabetic macular edema, 19 cases(19 eyes)of wet age-related macular degeneration and 25 cases(25 eyes)of retinal vein occlusion. Corneal confocal microscope was used to collect images of corneal subbasal nerve plexus before injections and at 1mo after each intravitreal injection based on 3+pro re nata(PRN)treatment regimen. Furthermore, the length and density of corneal nerve were measured.RESULTS: There was no significant difference in corneal nerve density of patients injected with aflibercept between pre-injection and post-injection(P>0.05), while the corneal nerve length after 2nd and 3rd injections was lower than that of pre-injection(all P<0.01). There were no significant changes in corneal nerve density and length in patients with intravitreal injections of ranibizumab(all P>0.05), and there was no significant differences in corneal nerve density and length after 3 injections of the two drugs(all P>0.05).CONCLUSION: Repeated intravitreal anti-VEGF drug may affect corneal nerve to some extent. For patients who need repeated intravitreal injections of anti-VEGF, attention should be paid to the changes of corneal nerves.

Integrating 16S sequencing and metabolomics study on anti-rheumatic mechanisms against collagen-induced arthritis of Wantong Jingu Tablet / 中国天然药物

Wantong Jingu Tablet (WJT), a mixture of traditional Chinese medicine, was reported to relieve the symptoms of rheumatoid arthritis (RA), but its pharmacological mechanism was not completely understood. The aim of this study was to investigate the therapeutic mechanisms of WJT for RA in vivo. The effects of WJT on joint pathology, as well as the levels of Bax, Bcl-2, caspase-3, cleaved-caspase-3, ERK1/2, pERK1/2, TNF-α, IL-1β, and IL-6 were measured using collagen-induced arthritis (CIA) rats. The intestinal flora composition and the metabolites alteration were analyzed by 16S rDNA sequencing and metabolomics method, respectively. We found that WJT ameliorated the severity of the CIA rats which might be mediated by inducing apoptosis, inactivating the MEK/ERK signals and reducing the production of pro-inflammatory cytokines. WJT, in part, relieved the gut microbiota dysbiosis, especially bacterial phylum Bacteroidetes, Tenericutes and Deferribacteres, as well as bacterial genus Vibrio, Macrococcus and Vagococcus. 3'-N-debenzoyl-2'-deoxytaxol, tubulysin B, and magnoline were significantly associated with the specific genera. We identified serotonin, glutathione disulfide, N-acetylneuraminic acid, naphthalene and thromboxane B2 as targeted molecules via metabolomics. Our findings contributed to the understanding of RA pathogenesis, and WJT played essential roles in gut microbiota health and metabolite modulation in the CIA rats.

Factors Related to Penile Erectile Function for Men with Spinal Cord Injury / 中国康复理论与实践

Objective:To analyze the clinical features related to erectile dysfunction for male patients with spinal cord injury. Methods:From December, 2019 to January, 2021, 28 male patients with spinal cord injury were detected the stiffness and periactivity index of penises during night sleep using RigiScan. The patients were grouped as presence or absence of bulbocavernous reflex, anal autonomic contraction, anal tactile sensation and anal pressure sensation, to compare the stiffness and periactivity index between the groups. Results:The stiffness and periactivity indexes were more in patients presenting bulbocavernosus reflex, anal autonomic contraction, anal tactile sensation and anal deep pressure sensation (|t| > 2.19, P < 0.05). Conclusion:Bulbocavernous reflex, anal autonomic contraction, anal tactile and anal deep pressure response may predict penile erectile function after spinal cord injury.

Strategy study of improving residents′ health literacy under the guidance of "Healthy Shanghai Action" / 上海预防医学

Objective Based on the Health Shanghai Action 2019-2030, combined with the monitoring results of Shanghai residents′ health literacy, we analyzed the data, looked for the existing problems and the possible causes in the improvement of health literacy, and put forward corresponding suggestions and strategies. Methods Using the monitoring data and results of Shanghai residents′ health literacy from 2008 to 2018, we made aggregate analysis of residents′ health literacy and comparative analysis of different years, different ages and different regions in this regard. Results The health literacy level of Shanghai residents was 28.38% in 2018.The absolute value of growth is 21.41% and the average annual growth rate was 13.61% compared with 2008.The literacy levels of infectious disease prevention, healthy behavior, and basic medical treatment were relatively low.There had been a big gap between urban and rural residents in literacy levels of health information, health knowledge, and health skill. Conclusion The health literacy level of Shanghai residents has been improved significantly.In future, we should strengthen the health education of infectious disease prevention, healthy behavior, and basic medical treatment, using different health education methods for different age groups or different education level people.Moreover, we need to establish and strengthen the authoritative health information release platform, so the residents especially rural ones, can get the reliable health education information in time.

Strategy study of improving residents′ health literacy under the guidance of "Healthy Shanghai Action" / 上海预防医学

Objective Based on the Health Shanghai Action 2019-2030, combined with the monitoring results of Shanghai residents′ health literacy, we analyzed the data, looked for the existing problems and the possible causes in the improvement of health literacy, and put forward corresponding suggestions and strategies. Methods Using the monitoring data and results of Shanghai residents′ health literacy from 2008 to 2018, we made aggregate analysis of residents′ health literacy and comparative analysis of different years, different ages and different regions in this regard. Results The health literacy level of Shanghai residents was 28.38% in 2018.The absolute value of growth is 21.41% and the average annual growth rate was 13.61% compared with 2008.The literacy levels of infectious disease prevention, healthy behavior, and basic medical treatment were relatively low.There had been a big gap between urban and rural residents in literacy levels of health information, health knowledge, and health skill. Conclusion The health literacy level of Shanghai residents has been improved significantly.In future, we should strengthen the health education of infectious disease prevention, healthy behavior, and basic medical treatment, using different health education methods for different age groups or different education level people.Moreover, we need to establish and strengthen the authoritative health information release platform, so the residents especially rural ones, can get the reliable health education information in time.

Association between serum levels of osteopontin and systolic pulmonary artery pressure among healthy adults post acute high altitude exposure / 中华心血管病杂志

Objective: To explore the association between serum levels of osteopontin (OPN) and systolic pulmonary artery pressure (sPAP) in healthy men following acute high altitude exposure. Methods: According to the inclusion and exclusion criteria, this observational study included 94 male subjects (aged from 18 to 30 years, dwelling in lowland<500 m) who ascended to Litang (4 100 m) from Chongqing (400 m) by bus with a stair-like journey within 7 days in June 2013. Data including basic information, OPN, superoxide dismutase (SOD), and malondialdehyde (MDA) and echocardiographic derived sPAP were collected within 48 hours before ascent and within 2-7 hours after arrival. Accordingly, subjects were divided into 3 groups based on the tertiles of sPAP after acute high altitude exposure: low sPAP group (26.8-32.3 mmHg (1 mmHg=0.133 kPa)) (n=31), middle sPAP group (32.4-37.4 mmHg) (n=32) and high sPAP group (37.5-55.6 mmHg) (n=31). Associations of serum OPN and SOD levels with sPAP were analysed by univariate and multivariate linear regression analysis. Results: After acute high altitude exposure, the levels of sPAP were significantly increased (P<0.001). There were no differences in age, height, weight, body mass index, percent of Han nationality and smoking among 3 subgroups. However, following acute high altitude exposure, the levels of heart rate, systolic and diastolic blood pressure elevated (all P<0.05), whereas the levels of oxygen saturation were reduced in the total subjects and all subgroups (all P<0.05). Moreover, systolic blood pressure of subjects in the high sPAP group was higher than that in low and middle sPAP groups (both P<0.05), and diastolic blood pressure of subjects in high sPAP group was higher than that in low sPAP group (P<0.05). The serum levels of OPN were increased in total cohort(27.9 (22.5,34.0) μg/L vs. 25.6 (18.4, 33.1) μg/L, P<0.05)， and high sPAP group (P<0.05), whereas no differences were found in serum SOD and MDA levels among groups. Furthermore, the serum level of OPN in high sPAP group was higher than that in low sPAP group at high altitude (P<0.05), and there was a trend for decline in SOD level with increasing sPAP (P>0.05). Results from univariable linear regression analysis showed that the serum levels of OPN (r=0.32, P=0.002) and SOD (r=-0.22，P=0.032) were linearly correlated with sPAP in total cohort after high altitude exposure. Multivariate regression analysis showed that the serum levels of OPN(β=0.310，P=0.002) and SOD (β=-0.199，P=0.043) were independently associated with the levels of sPAP at high altitude. Conclusion: After acute high altitude exposure, the serum level of OPN is positively associated with sPAP, suggesting that OPN may be a novel bio-marker for predicting the increase of pulmonary pressure in response to acute high altitude exposure.

Construction of swept source optical coherence tomography imaging system for root canal endoscopy and application in diagnosis of root fractures / 北京大学学报(医学版)

OBJECTIVE@#To construct swept source optical coherence tomography (SS-OCT) imaging system for root canal endoscopy, and to evaluate the accuracy of diagnosing artificial root fractures.@*METHODS@#An ultra-high-speed (40 kHz) swept laser source was developed based on the piezoelectric tuning filter and the Fourier domain mode locking (FDML) swept laser technology (patent number: 200620135940.2). Ultra-miniature gradient index lens technology (patent number: 201320241218.7) was used to create a thin endoscopic probe with a diameter of 0.86 mm for real-time image transmission. The SS-OCT light source had a wavelength of 1 310 nm and a bandwidth of 100 nm. The axial and transverse image resolutions were 15 μm and 25 μm, respectively. Artificial fractures were created on human mandibular premolars with single root and the premolar roots were prepared to 41 horizontal sections (1 mm thick). 27 root sections with fractures (width: 52-284 μm) and 14 the sections without fractures were observed under an optical stereomicroscope with a cold light source as the gold standard. The horizontal root sections were scanned by self-developed SS-OCT imaging system for root canal endoscopy with a central wavelength of 1 310 nm and bandwidth of 100 nm. The data were reconstructed with 30 μm thick slices at an interval of 30 μm. Two observers, a radiologist and an endodontist, were trained and independently evaluated all the reconstructed images blindly. The diagnostic performance of SS-OCT imaging system was calculated, and statistical analysis was performed.@*RESULTS@#SS-OCT root canal endoscopic imaging system composed of high-speed swept laser source, fiber coupler, endoscopic probe, reference arm and differential detector. Root sections could be scanned by SS-OCT and imaged in realtime at a depth of 1 to 2 mm. The kappa value for interobserver agreement was 0.792, and the intraobserver agreement was 1.000 and 0.709 for two observers respectively. All of 27 fractured root sections and 12 of 14 root sections without fractures were accurately diagnosed while 2 unfractured root sections were misdiagnosed. The sensitivity was 1.000 and the specificity was 0.857 for diagnosis of artificial root fractures by SS-OCT. The positive predictive value (PPV), negative predictive value (NPV) and the overall accuracy rate were 0.931, 1.000 and 0.951 respectively.@*CONCLUSION@#The swept source optical coherence tomography imaging system for root canal endoscopy is a promising imaging method for observing root fractures．.

Study on expression of long non-coding RNA in colon cancer tissues and adjacent tissues / 中国病理生理杂志

AIM:To screen the differentially expressed long non-coding RNA(lncRNA)in colon cancer,and to explore its expression in colon cancer tissues and adjacent tissues.METHODS:The "Colon adenocarcinoma:Person neoplasm cancer status" which consisted of 36 cases of colon cancer tissues and 29 cases of normal colonic tissues was downloaded from the lncRNAtor database.The candidate genes were selected from these differentially expressed lncRNAs based on artificial criterion(P<0.01;fold change ≥2 or<0.5)and then validated by real-time PCR in 60 pairs of colon cancer tissues and adjacent tissues.RESULTS:A total of 50 lncRNAs were differentially expressed in colon cancer tis-sues,including 28 up-regulated and 22 down-regulated(P<0.01).The verifying results displayed that HNF1A-AS1 and ZDHHC8P1 were up-regulated(P<0.01),and SUZ12P expression was down-regulated(P<0.05),but the expression of AC069513.3 was not statistically significant between colon cancer tissues and adjacent tissues.The abilities of HNF1A-AS1,ZDHHC8P1,SUZ12P and AC069513.3 to discriminate the colon cancer from normal adjacent tissue by the ROC curve with an AUC of 0.729(sensitivity 78％,specificity 67％),0.617(sensitivity 68％,specificity 55％),0.689(sensitivity 66％,specificity 55％)and 0.518(sensitivity 52％,specificity 48％)were observed.CONCLUSION:Long non-coding RNA HNF1A-AS1 and ZDHHC8P1 are up-regulated and SUZ12P is down-regulated in colon cancer tis-sues,suggesting that they may be involved in the pathogenesis of colon cancer.

Detection of root cracks after root canal preparation using rotary NiTi systems by optical coherence tomography (OCT) scan / 北京大学学报(医学版)

OBJECTIVE@#To evaluate the accuracy of optical coherence tomography (OCT) in detecting root cracks after root canal instrumentation using histological gold standard.@*METHODS@#Twenty complete extracted human mandibular incisors that were free of caries, calculus, and root treatment were chosen and accessed coronally with a diamond bur, then mounted in resin blocks with alginate impression material using simulated periodontal ligaments, and the apex was exposed 3 mm. The teeth were stored in water at room temperature. Then the teeth were then instrumented to the major apical foramen (AF) at #30/0.09 using ProTaper Universal rotary nickel titanium system (Dentsply Maillefer, Ballaigues, Switzerland), irrigation with 1% sodium hypochlorite (NaOCl) by using a 26-gauge needle followed after each instrument. The apical root was scanned with 360° of rotation by Swept-Source Optical Coherence Tomography (SS-OCT) (wavelength: 1 310 nm, scan rate: 20 kHz, axial resolution: 16 μm) with driving device (stepper motor and lifting platform). The reconstruction images of axial planes 1, 2 and 3 mm from the apex were examined and the root cracks were blindly diagnosed by two observers. The horizontal section was performed at 1, 2 and 3 mm from the apex using low speed disc saw (Leica SP1600, Wetzlar, Germany). The presence of cracks was noted under an optical stereomicroscope (ZOOM-630E) with a cold light source using as gold standard to evaluate the accuracy of OCT in detecting root cracks after root canal instrumentation.@*RESULTS@#After canals instrumentation with ProTaper Universal rotary nickel titanium system to #30/0.09, root cracks were detected in 9 of 20 teeth by histological examination. Crack lines were observed on 13 of 60 horizontal sections and cracks on 12 of the 13 sections were detected by OCT. No cracks were observed in the other 47 of the 60 horizontal sections,none of which was misdiagnosed by OCT. The overall accuracy rate for detection of root cracks with OCT was 0.983, the sensitivity was 0.923, the specificity was 1.000, the positive predictive value (PPV) was 1.000 and the negative predictive value (NPV) was 0.979.@*CONCLUSION@#OCT may be a promising nondestructive imaging method for diagnosing root canal cracks after canal instrumentation.

Metabolism evaluation of blood quality after marine storage and transportation / 军事医学

Objective To evaluate the quality of blood after marine storage and transportation,to establish a quantitative energy metabolism platform supported by extracellular flux(XF)technique to determine the glycolysis level of red blood cells(RBC), and to introduce the "functional dose", as a valuable supplement, into the quality evaluation of RBCs after marine storage and transportation.Methods The extracellular acidification rate(ECAR)of erythrocyte glycolysis was detected by XF.The levels of glycolysis, ATP contents and free hemoglobin of RBCs were detected before and after marine navigation.RBC suspensions stored at 4℃in a blood bank on land served as control.Results The results showed that the glycolysis level of RBCs was detected by XF.Indicators of blood quality and energy metabolism of RBCs were detected in the twice marine storage and transportation.Blood samples transported around the Gulf of Bohai had no significant change,indicating that the samples were well preserved.The energy indicators of blood samples navigated in the Pacific Ocean decreased significantly,and it was recommended that the samples be discarded.Conclusion XF technique has been applied to the measurement of erythrocyte glycolysis,enabling high-throughput and multi-indicator dynamic detection of viable cells.The energy metabolism of erythrocyte should be recommended as an important indicator for evaluating blood after marine storage and transportation.

Identification of C C Location of Unsaturated Phosphatidylcholines in Cell by Photochemical Reaction-Tandem Mass Spectrometry / 分析化学

A method of identification of C=C location and relative quantitation of unsaturated phosphatidylcholine ( PC ) isomers in breast cells by online photochemical reaction-pulsed directed current electrospray-tandem mass spectrometry ( PB-pulsed-dc-ESI-MS/MS) was established with benzophenone ( BP) as a photochemical reactant. The three-phase extraction method was used to extract the lipids in the cells, and then the C=C in the unsaturated PC and the carbonyl in BP were specifically cycled under the irradiation of 254 nm ultraviolet light (Paternò-Büchi, PB reaction). The PB products were ionized and mass-isolated for low-energy collision dissociation through the non-contact pulsed-dc-ESI ionization method. The double bond position and the relative content of the location isomers were obtained from the resulting ions in the MS/MS spectrum. The C=C location of 8 kinds of unsaturated PCs in MCF-7 and MCF-10A was detected, and the relative contents of 4 kinds of C=C location isoforms were analyzed. It was found that the relative abundance of △9 isomer in PC 16:018:1 was not significantly different between the two cells. The relative abundance of△9 isomers in PC 18:018:1 and PC 18:118:1 was slightly different. However, there is a big difference of △9 in LPC 18:1 between the cancer cell and normal cell (56. 0% ± 1. 3% vs. 71. 7% ± 6. 8%). The establishment of such a rapid and easy mass spectrometry method can analyze the C=C location and the relative content of location isomers, and it is expected to be a powerful tool to identify different cell states and different disease states.

Clinical features of late-onset neuromyelitis optica spectrum disorders / 天津医药

Objective To explore the clinical features of late-onset neuromyelitis optica spectrum disorders (LON?MOSD). Methods A retrospective analysis was performed to evaluate 61 patients with LONMOSD admitted to our hospital from January 2010 to May 2015. Results (1) The median age at onset was 57 (53, 63) years, male/female was 1∶3.7. Thirty-two patients (52.5%) had transverse myelitis (TM) and 16 patients (26.2%) had optic neuritis (ON) at the disease onset. Fifty-one patients (83.6%) experienced recurrent attacks. Forty patients (65.6%) showed abnormal brain magnetic resonance imag?ing (MRI). Spinal cord MRI showed more frequently present in thoracic regions (39.3%). (2) There were no significant differ?ences in clinical features between AQP-4 seropositive and seronegative groups. (3) By Spearman analysis, it was obvious that EDSS scores at acute phase and remission were positively correlated to AQP-4 antibody levels (rs=0.389, P0.05;rs=0.096, P>0.05). Conclusion LONMOSD patients are more prone to present with TM at onset and have more lesions in thoracic spinal cord and brain. The AQP-4 antibody titres can indicate the severity of disease in acute phase.

Expansion of chicken bone marrow mesenchymal stem cells by laminin culture system / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cel s from chickens are important cel models for embryonic developmental biology, immunology and oncology research. However, it is difficult to keep bone marrow mesenchymal stem cel s with good undifferentiated potential in a large-scale expansion system. OBJECTIVE:To establish a culture system in vitro with laminin coating to expand bone marrow mesenchymal stem cel s from chickens. METHODS:Isolated bone marrow mesenchymal stem cel s from chickens were seeded in laminin-coated plates and traditional two-dimensional plates, respectively. After expansion in vitro, the morphological characteristics, expression of surface markers, expansion characteristics and adipogenic differentiation of bone marrow mesenchymal stem cel s in both conditions were analyzed and compared. RESULTS AND CONCLUSION:There were no statistical differences in the morphological characteristics and expression of surface markers of bone marrow mesenchymal stem cel s expanded by laminin-coated plates and traditional two-dimensional plates. But, the expansion characteristics and adipogenic differentiation of bone marrow mesenchymal stem cel s cultured in laminin-coated plates were better than those in traditional two-dimensional plates. Laminin culture system could quickly amplify out of a large number of chicken bone marrow mesenchymal stem cel s with better proliferation ability and undifferentiated performance. Al above results indicated that a more efficient expansion system with laminin coating is established.

Biological characteristics and superiority of rat bone marrow mesenchymal stem cells isolated and cultured using whole bone marrow adherence method / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cells are rare in vivo. It is important to purify, proliferate and differentiate bone marrow mesenchymal stem cells in vitro for further research. OBJECTIVE:To evaluate the biological characteristics, phenotype and multiple differentiation potential cultivation of bone marrow mesenchymal stem cells that are isolated, cultured and purified using the whole bone marrow adherence method. METHODS:Bone marrow mesenchymal stem cells were isolated, purified and cultured by the whole bone marrow adherence method. Morphological observation and flow cytometry determination of cellsurface markers were performed. Osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells was induced. RESULTS AND CONCLUSION:We successful y purified and proliferated bone marrow mesenchymal stem cells with high cellviability and differentiation ability. Fibroblast-like cells were harvested, expressing CD29 and CD90, but not CD45. Fol owing osteogenic and adipogenic induction, cells were positive for oil red O staining and alizarin red staining. The whole bone marrow adherence method is easy to operate, has little impact on cellviability, and can be used to harvest high-purification bone marrow mesenchymal stem cells with high cellviability and differentiation ability.

Analysis of the efficacy and prognosis on first-line autologous hematopoietic stem cell transplantation of patients with multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the efficacy and prognosis of first-line autologous hematopoietic stem cell transplantation (ASCT) for newly diagnosed patients with multiple myeloma(MM).</p><p><b>METHODS</b>From January 2005 to December 31, 2012, 60 patients with MM were enrolled. All patients received thalidomide or/and bortezomib-based induction therapy, then received high-dose melphalan (200 mg/m²) and autologous stem cell support to get a ≥ partial response (PR), and followed by thalidomide-dexamethasone (TD) ±bortezomib as consolidation or maintenance treatment. With the follow up to December 31, 2012, the overall survival (OS), progression free survival (PFS) and the prognostic factors, including ISS stage, response and fluorescent in situ hybridization (FISH) data of cytogenetics were analyzed.</p><p><b>RESULTS</b>With a median follow up of 36.8 (12.0-102.5) months, the median OS and PFS estimate were not reached and 86.5 months, respectively. After transplantation, all (100%) patients received very good partial response (VGPR), and 34 (56.7%) patients achieved complete response (CR) after consolidation or maintenance treatment. The patients that achieved CR resulted in long term PFS (P=0.030), with no difference in OS (P=0.942). The univariate analysis showed that the abnormalities, including 13q14 deletion, 1q21 gain, IgH location and p53 deletion had the prognostic impacts. If the t(4;14) or p53 deletion was excluded, there would be no correlation between 13q14 deletion or 1q21 gain with PFS and OS. The patients with p53 deletion had a worst survival.</p><p><b>CONCLUSION</b>There has been significant improvement in the outcome for young MM patients by using ASCT and novel drugs. Cytogenetic abnormalities and response to therapy are the main factors affecting the survival of patients.</p>

Clinical efficacy and safety of chemoimmunotherapy with rituximab,fludarabine and cyclophosphamide for chronic lymphocytic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of a chemoimmunotherapy regimen of rituximab, fludarabine and cyclophosphamide (FCR) for patients with chronic lymphocytic leukemia(CLL).</p><p><b>METHODS</b>The clinical data of 26 CLL patients receiving FCR regimen in our hospital from April 2003 to January 2012 were analyzed retrospectively. Patients were grouped according to indicators including Rai risk stratification, β(2)-MG, LDH, ZAP-70, CD38, cytogenetics and immunoglobulin heavy chain variable region gene (IgVH) mutation status. Therapy efficacy and survival were evaluated and the safety of FCR regimen was assessed.</p><p><b>RESULTS</b>Among 26 patients, the overall response rate ( ORR ) was 76.9%, 10 patients (38.5%) achieved complete remission(CR) and 10(38.5%) partial remission(PR). With a median follow-up time of 30 ( 3-98 ) months, the median estimated progression-free survival(PFS) for all patients was 42(16-68) months and median overall survival(OS) was 63(41-85)months. Clinical parameters associated with higher CR rates were ＜2 courses of prior treatment regimens, proportions of bone marrow lymphocytes declining ≥ 50% after 2 courses of FCR, low LDH, low β(2)-MG and ZAP-70 negative (P = 0.014, 0.008, 0.027, 0.035 and 0.013, retrospectively). PFS and OS time in minimal residual disease(MRD)-negative, normal LDH and proportions of bone marrow lymphocytes declining ≥ 50% after 2 courses of FCR patients were significantly better than that of the control group (P＜0.05), PFS in the non-high-risk genetics group was significantly better than that in the high-risk genetics group (P = 0.005), while OS between two groups showed no statistically significant difference. The most common toxicities were gastrointestinal reactions (88.5%), followed by bone marrow suppression (80.8%): including neutropenia, anemia and thrombocytopenia. Infections accounted for 30.8%, mainly lung infection.</p><p><b>CONCLUSION</b>FCR is an effective and well-tolerated therapy for patients with CLL. Patients with MRD-positive, elevated LDH, proportions of bone marrow lymphocytes declining＜50% after 2 courses of FCR and high risk genetics patients are suitable for more effective treatment after achieving treatment response.</p>

Outcomes of younger than 60 years old adults with Ph/BCR-ABL positive acute lymphoblastic leukemia: a single center clinical trial of BDH ALL 2000/02 / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the treatment options for younger than 60 years old adults with Ph /BCR-ABL positive acute lymphoblastic leukemia (Ph⁺ ALL).</p><p><b>METHODS</b>From January 2001 to June 2012, 42 adult patients were enrolled in the study. All patients received standard VDCP±L ±imatinb (IM) as induction therapy followed by intensive consolidation of modified Hyper-CVAD/MA±IM. At complete remission 1 (CR1), patients with appropriate donor received allogeneic hematopoietic stem cell transplantation (allo-HSCT), the others sequentially received intensive consolidation ±IM and autologous HSCT (ASCT) at molecular CR (MCR), then MM±VP±IM as maintenance therapy. Overall survival (OS), disease free survival (DFS) and relapse rate (RR) were analyzed.</p><p><b>RESULTS</b>CR rate after 1 cycle of induction chemotherapy was 83.3%. 39(92.9%) patients achieved CR. The median DFS and OS were (22.0±3.5) and (37.0±5.3) months respectively, with cumulative RR of (43.7±9.7)% during a median follow-up of 26.5(8-75) months. All 7 patients in CT group relapsed. Two patients received IM pre- and post-ASCT maintained MCR for 35 and 12 months after ASCT. But the other 3 ASCT recipients without IM died of relapse within 1 year. The transplant-related mortality rate in allo-HSCT group was 12.5%. The estimated 3-year OS in allo-HSCT (n=16), ASCT (n=5) and CT (n=7) groups were (66.7±12.2)%, (25.0±21.7)% and (16.7±15.2)%, respectively (P=0.014); meanwhile, the estimated 3-year DFS in those groups were of (56.3±12.4)%, (26.7±22.6)% and 0, respectively (P=0.002).</p><p><b>CONCLUSION</b>IM combined with intensive chemotherapy significantly increased the CR rate with the improved quality of CR, which highlighted the feasibility of SCT. Allo-HSCT could decrease relapse to produce favorable OS and DFS in CR1 of young adults with Ph⁺ ALL. ASCT combined IM might be the treatment of choice for those achieved MCR but without donors.</p>