Postoperative imatinib treatment in gastric intermediate-risk gastrointestinal stromal tumors / 中华普通外科杂志

Objective To explore the clinical prognosis and efficacy of adjuvant therapy with imatinib of postoperative patients with gastric intermediate-risk gastrointestinal stromal tumor (GIST).Methods The clinicopathological data and follow-up data of 93 gastric intermediate-risk GIST cases from Jan 2005 to Dec 2016 at Union Hospital were analyzed retrospectively.Univariate and multivariate analysis were performed to assess the prognostic factors.Results There were 93 patients undergoing complete GIST resection with 42(45％) cases receiving post-op imatinib 400 mg/d for targeted therapy.The median target therapy period was 12 (6-72) months.86％ (80 cases) patients were followed up for 46 (6-120) months.The 1-,3-,5-year recurrence-free survival rate (RFS) of the whole group were 100％,91.5％,88.5％ respectively.Multivariate analysis revealed that mitotic count (P =0.040,RR =6.078,95％ CI:0.541-68.274) and neutrophil-lymphocyte ratio (NLR) (P =0.036,RR =6.102,95％ CI:0.782-47.632) were prognostic risk factors of RFS.For those mitotic count ＞ 2/50 HPF and NLR ＞ 2.3,adjuvant therapy with imatinib significantly increases RFS.Conclusion Mitotic count and NLR were independent risk factors of RFS in gastric intermediate-risk GIST.For those with mitotic count ＞ 2/50 HPF and NLR ＞ 2.3,postoperative adjuvant therapy with imatinib helps improve the prognosis.

Immunotherapy for acute myeloid leukemia: how many achievements can be converted into clinical practice? / 白血病·淋巴瘤

Acute myeloid leukemia (AML) is a highly heterogeneous hematologic malignancy with a high relapse rate. In recent years, immunotherapy has achieved certain curative effects in AML. A considerable proportion of AML patients can achieve long-term survival after allogeneic hematopoietic stem cell transplantation, adoptive immunotherapy, monoclonal antibody and vaccine therapy. But immunotherapy still can not meet the needs of all AML patients. Immunotherapy of AML has great potential for development, and it is expected that various immune therapies can better improve the efficacy of AML in the case of giving full play to their advantages and reducing side effects.

Experimental study on expression of TH and GDNF bi-gene of lentivirus by improved Tet-On system for a rat model of Parkinson’ s disease / 中国药理学通报

Aim To investigate TH and GDNF genes expression and regulation of lentivirus ( Lv-TH-GDNF ) based on improved Tet-On system and the effect of the Lv-TH-GDNF intrastriatal transfer on a rat Parkinson’ s disease( PD) model. Methods 1. HeLa cells were infected by obtained Lv-TH-GDNF and rtTA2 s-M2 vi-rus. The expression of tyrosine hydroxylase ( TH ) and glial cell line-derived neurotrophic factor( GDNF) genes was induced by doxycycline( Dox) which was examined by Western blot. 2. The Lv-TH-GDNF together with rtTA2 s-M2 viruses were injected into lesion-side stria-tum of a rat PD model, and the expression of GDNF and TH genes was induced by Dox. Then, the effects of Lv-TH-GDNF were evaluated by the apomorphine-induced rotational behavior, the number of dopaminer-gic neurons in substantia nigra,DA and DOPAC levels in the lesion-side striatum. In addition, Western blot was performed to check the expression of TH and GD-NF genes in the transplanted striatum. Results 1. In vitro studies on HeLa cells, Western blot showed clear protein bands of TH and GDNF in the Dox-positive group, but not in the Dox-negative group. 2. In vivo experiments in animals, the results showed that, 4 weeks after transplantation, the apomorphine-induced turning effect was significantly improved ( P<0 . 01 ) , the number of TH-positive cells in the lesion-side sub-stantia nigra pars compacta as well as the content of DA and DOPAC, the protein level of GDNF and TH genes in the lesion-side striatum was significantly in-creased ( P<0 . 01 ) , each of which was only in Lv-TH-GDNF+rtTA2 s-M2+Dox-treated rats as compared with PBS-treated rats. Conclusion The expression of TH and GDNF genes in Lv-TH-GDNF based on im-proved Tet-On system is effectively regulated by tetra-cycline antibiotics without basal activity in vitro, and the intrastriatal transfer of which has certain therapeutic effect on PD rats.

New progress of adoptive immunotherapy in the treatment of hematologic malignancies based on chimeric antigen receptor T-cells: reports from the 56th American Society of Hematology annual meeting / 白血病·淋巴瘤

T cells redirected to specific antigen targets with engineered chimeric antigen receptors (CARs) are emerging as powerful therapies in hematologic malignancies.Various CAR designs,manufacturing processes,and study populations,among other variables,have been studied and reported in clinical trials.The accumulating data supporting CAR-T cells therapy for disease such as chronic lymphocytic leukemia (CLL)and acute lymphocytic leukemia (ALL) highlight what may well become the next sea change in the care of patients with all types of hematologic neoplasia.The scientific symposium on CAR-T cells therapy inspires the mounting enthusiasm regarding this new treatment strategy.Here,the results of the reported clinical trials and the outlook for CAR-T cells therapies were reviewed and discussed.Many questions remain in the field of CAR-T cells directed to hematologic malignancies,but the encouraging response rates pave a wide road for future investigation.

Expression of A20 in patients with systemic lupus erythematosus / 中国病理生理杂志

[ ABSTRACT] AIM:To investigate the expression and regulation of A20 in healthy individuals and the patients with systemic lupus erythematosus (SLE).METHODS:The expression levels of A20, NF-κB, MALT1, and MALT1V1 in peripheral blood mononuclear cells ( PBMC) of the patients with SLE ( including 2 cases with scleroderma, 1 case with rheumatoid arthritis, and 1 case with lymphoma) were analyzed by real-time PCR.RESULTS:A significantly lower A20 expression level was found in the PBMC from SLE group compared with the healthy controls, while the expression levels of MALT1 and NF-κB were also decreased.In addition, no significant correlation between A20 and NF-κB expression levels in healthy group was observed, but a positive correlation was found in SLE group ( P<0.05) .A significant positive corre-lation between MALT1 and NF-κB expression levels in healthy group ( P<0.05) was observed, and no significant correla-tion was found in SLE group.The expression level of MALT1V1 in SLE group was significantly lower than that in healthy control group, and there was a positive correlation between A20 and MALT1V1 in healthy volunteers (P<0.01), but that did not exist in SLE group.CONCLUSION: The characteristics of the expression pattern of MALT1-A20-NF-κB in the SLE patients were presented.Lower level of A20 expression was found in the SLE patients, in particular with other autoim-mune disease or lymphomas, indicating the lower immune tolerance in SLE.The positive correlation of A20 and NF-κB may relate to positive regulation of MALT1.

Establishment and evaluation of detection method of bifidobacteria in human fecal using real-time fluorescence quantitative PCR / 吉林大学学报(医学版)

Objective To establish the real-time fluorescence quantitative PCR method for the detection of bifidobacteria in human fecal samples, and to provide an effective means for measuring intestinal bacteria. Methods Total DNA of bacteria was extracted from 60 cases of children's fecal samples. Three primers of bifidobacteria based on the 16S ribosomal RNA (16SrRNA)which possessed specialities of bacteria as amplified region were designed.The part of amplified 16SrRNA gene sequences was used as standard production.The serial dilution of standard was analyzed to build an absolute quantitative standard curve with SYBR GreenⅠ dye method, and the bifidobacterium contents in sixty human fecal samples were calculated. The sensitivity of the reaction was calculated by detecting the lowest detectable standard which determined the sensitivity of the reaction. The PCR products’melting curve was used to evaluate the specificity.The coefficient of variation (CV)of different batches of standard with the same concentration was used to evaluate the stability of reaction.Results The length of PCR product fragment which was used to build the standard curve was about 6 1 3 bp, the sequencing result was consist with the goals, and the standard sample of bifidobacteria was successfully established in real-time fluorescence quantitative PCR.The standard curve showed a good linear relationship with R2=0.999.The minimum detection value was 1.48×102 copies per reaction.The melting curve of real-time fluorescence quantitative PCR was a single peak.The test samples were batched and then examined by fluorescence quantitative PCR.The CV of standards’ Ct values which calculated from the standard (1.48 × 103 -1.48 × 107 copies · μL-1 )were 2.94%, 3.39%, 3.54%,3.08%,and 3.34%,respectively.The contents of bifidobacteria in fecal from 60 children was 7.77± 0.86(copies · g-1 wet fecal)transformed by logarithmic.Conclusion The established real-time fluorescence quantitative PCR method has high sensitivity, strong specificity and good repeatability, which is suitable for detection of human fecal bifidobacteria content.

Non-anthraquinones constituents from the roots of Knoxia valerianoides / 中国中药杂志

Twenty-one non-anthraquinones constituents were isolated for the first time from an ethanol extract of the roots of Knoxia valerianoides by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were identified by their physical-chemical properties and spectroscopic analysis including NMR and MS. The compounds include ten triterpenoids: ursolic acid (1), oleanolic acid (2), 2-oxo pomolic acid (3), pomolic acid (4), maslinic acid (5), rotungenic acid (6), tormentic accid (7), rotundic acid 3,23-acetonide (8), arjungenin (9), and 2alpha, 3beta, 19alpha, 23-tetrahydroxy-urs-12-en-28-oic acid (10), four sitosterones: (24R)-24-ethylcholesta-4,22-dien-3-one (11), 3-oxo-4-en-sitosterone (12), 7-oxostigmasterol (13), and 7-oxo-beta-sitosterol (14), two lignans: eudesmin (15) and ciwujiatone (16), one coumarin: cnidilin (17), and four simple aromatic analogues: 5-hydroxymethylenefural (18), 3-hydroxy-4-methoxybenzoic acid (19), benzoic acid (20), and 2-hydroxy-5-methxoycinnamaldehydes (21). In the in vitro assays against human cancer cell lines (HCT-8, Bel7402, BGC-823, A549, and A2780), against deserum and glutamate induced PC12-syn cell damage, and against HIV-1 replication, and inhibiting protein tyrosine phosphatase 1B (PTP1 B), LPS induced NO production in macrophage, and Fe(2+)-cystine induced rat liver microsomal lipid peroxidation, at a concentration of 1 x 10(-5) mol x L(-1), no compound showed activity.

Chemical constituents from roots of Machilus yaoshansis / 中国中药杂志

To study chemical constituents contained in ethanol extracts from roots of Machilus yaoshansis. Fifteen compounds were separated from the roots of M. yaoshansis by using various chromatographic techniques. Their structures were identified on the basis of their physicochemical properties and spectral data as twelve lignans(+)-guaiacin (1), kadsuralignan C (2), (+)-isolariciresinol (3), 5'-methoxy-(+)-isolariciresinol (4), (7'S, 8R, 8'R)-lyoniresinol (5), meso-secoisolariciresinol (6), isolariciresinol-9'-O-beta-D-xylopyranoside (7), 5'-methoxy-isolariciresinol-9'-O-beta-D-xylopyranoside (8), lyoniresinol-9'-O-beta-D-xylopyranoside (9), (2R, 3R) -2, 3-dihydro-2-(4-hydroxy-3-methoxyphenyl)-7-methoxy-3-methyl-5-(E)-propenylbenzofuran (10), 3, 5'-dimethoxy-4', 7-epoxy-8, 3'-neolignan-4, 9, 9'-triol (11), nectandrin B (12), and three flavanes(+)-catechin (13), (-)-epicatechin (14), and bis-8, 8'-catechinylmethane (15). All of the compounds 1-15 were separated from M. yaoshansis for the first time.

Studies on constituents of cultures of fungus Phellinus igniarius / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of the culture of Phellinus igniarius and their phamacological activities.</p><p><b>METHOD</b>The constituents were isolated by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Structures of the isolates were identified by spectroscopic data analysis. Cytotoxic, neuroprotective, hepatoprotective, anti-inflammatory, and anti-HIV activities were screened by using cell-based models.</p><p><b>RESULT</b>Twenty-nine constituents were isolated. Their structures were identified as three sesquiterpenes: 3S,9R,10S-3-hydroxy-11, 12-O-isopropyldrimene(1), 3S, 9R, 10S-3, 11, 12-trihydroxydrimene (2), and 3S, 4S, 9R, 10S-11, 12, 14-trihydroxydrimene(3); three steriods: 24R-ergosta-4, 6, 8(14), 22-tetraen-3-one (4), stigmasta-7, 22-diene-3b, 5a, 6a-triol (5), and 5a, 8a-epi dioxyergosta-6, 22-diene-3b-ol (6); fourteen cyclo-dipeptide: cyclo (L-Pro-L-Val) (7), cycle (L-Leu-D-Pro) (8), cyclo (L-Leu-L-Pro) (9), cyclo (ILe-Pro) (10), cyclo (Gly-Leu) (11), cyclo (Phe-Ser) (12), cyclo (Ala-Pro) (13), cyclo (Ala-Phe) (14), cyclo (4-HyP-Phe) (15) , cyclo (L-Phe-D-Pro) (16), cyclo (D-Phe-D-Pro) (17), cyclo (6-HyP-Phe) (18), cycle (Gln-Pro) (19), and cycle (Asn-Leu) (20); and nine other compounds: N-acetyl-phenylalanine (21), adenosine (22), phenyldiethanol (23), o-hydroxy-phenylethanol (24), benzoic acid (25), p-methoxybenzoic acid (26), m-methoxybenzoic acid (27), hexadecanoic acid (28), and 3-pyridinecarboxylic acid (29). In the in vitro assays, at a concentration of 1 x 10(-5) mol x L(-1), compounds 5 and 8 showed neuroprotective activity against MPP+ induced PC12-syn cell damage, with a relative cell proliferation rate of 90.3% and 87.5% (P < 0.05). At 1 x 10(-5) mol x L(-1), compounds 12 and 18 showed hepatoprotective activities against DL-galactosamine-induced toxicity examined in WB-F344 cell, with cell survival rates of 25% and 24%, respectivily.</p><p><b>CONCLUSION</b>Compounds 1-29 were obtained from P. igniarius for the first time. Compounds 5 and 8 showed potent PC12-syn protective activities, while 12 and 18 showed hepato cytes (WB-F344 cells) protective activities.</p>

Anthraquinones from the roots of Knoxia valerianoides / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the chemical constituents of the roots of Knoxia valerianoides and their biological activities.</p><p><b>METHOD</b>The anthraquinones were isolated by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Structures of the isolates were identified by their physical-chemical properties and spectroscopic analysis including 2D NMR and MS. Antioxidant, anti-HIV, neuroprotective, and cytotoxic activities were screened by using cell-based models.</p><p><b>RESULT</b>Twenty-two constituents were isolated from an ethanolic extract of the roots of K. valerianoides. Their structures were identified as nordamnacanthal (1), ibericin (2), rubiadin (3), damnacanthol (4), 2-ethoxymethylknoxiavaledin (5), 3-hydroxymorindone (6), knoxiadin (7), 2-formyl knoxiavaledin (8), lucidin (9), xanthopurpurin (10), 1, 3-dihydroxy-2-methoxy-9, 10- anthraquinone (11), lucidin(-methyl ether (12), digiferruginol (13), 3-hydroxy-2-methyl-9,10-anthraquinone (14), rubiadin-1-methyl ether (15), 6-methoxylucidin (-ethyl ether (16), 1,3,6-trihydroxy-2-methyl-9,10-anthraquinone (17), 1,3-dihydroxy-2-hydroxy methyl-6-methoxy-9,10-anthraquinone (18), 1,3,6-trihydroxy-2-methoxymethyl-9,10- anthraquinone (19), 3,6-dihydroxy-2- hydroxymethyl-9,10-anthraquinone (20), and 1,6-dihydroxy-2-methyl-9,10-anthra quinone (21). In the in vitro assays, at a concentration of 1 x 10(-5) mol x L(-1), no compounds were active against human cancer cell lines (HCT-8, Bel7402, BGC-823, A549, and A2780), deserum and glutamate induced PC12-syn cell damage, LPS induced NO production in macrophage, Fe2+-cystine induced rat liver microsomal lipid peroxidation, HIV-1 replication, and protein tyrosine phosphatase 1B (PTP1B).</p><p><b>CONCLUSION</b>Compounds 9-21 were obtained from the roots of K. valerianoides for the first time.</p>

Overexpression of Nanog gene in mouse mesenchymal stem cells and its influence on NF-kappaB expression / 生物工程学报

The expression of NF-kappaB is considered to be involved in the progress of neurodegeneration. It has been reported that Nanog can suppress the expression of NF-kappaB. To inspect and verify this finding, we constructed lentivirus (LV) vector that overexpressed the Nanog gene, infected mouse mesenchymal stem cells (mMSCs), and examined the influence of Nanog overexpression on NF-kappaB gene expression. The plasmid pNL-Nanog-IRES2-EGFP was constructed by double digestion and genetic recombination. Sequencing results confirmed that our cloned Nanog gene in the PNL-Nanog-IRES2-EGFP plasmid was consistent with the sequence reported in the GenBank. The three plasmids: pNL-Nanog-IRES2-EGFP, HELPER, and VSVG were cotransfected into 293T cells to produce LV particles. After co-transfection of the three lentiviral plasmids, green fluorescence was observed confirming successful transfection. The mMSCs were infected by the LV and the expression of Nanog was then also verified by the presence of green fluorescence. Nanog expression levels in the mMSCs were examined using Western blotting. Expression of NF-kappaB was also examined using RT-PCR and Western blotting, and in addition with fluorescent microscope after immunocytochemical staining. The levels of Nanog protein expression in Nanog-mMSCs were significantly increased, and the levels of NF-kappaB mRNA and protein expression in Nanog-infected mMSCs were significantly lower than those of Mock-mMSCs and the mMSCs control groups. Our findings suggest that mMSCs genetically modified to overexpress Nanog can lead to the suppression of NF-kappaB expression. This suppression of NF-kappaB could have important implications for the treatment of neurodegeneration, and hence further scientific investigations of these interactions will have significant impact on future clinical attempts to attenuate disease progression.

Digital Molybdenum Target Mammography in Diagnosis of Breast Cancer / 实用放射学杂志

Objective To study the value of digital molybdenum target mammography in diagnosing breast cancer.Methods Digital molybdenum target X-ray findings of 36 cases with breast cancer confirmed by pathology were retrospectively analysed.Results X-ray appeared as masses in 22 cases(61.1%),masses with calcification in 7(19.4%),asymmetry increased density with structural disorder in 4(11.1%),Pagets disease in 1(2.7%),pure calcification in 3(8.33%),abnormal vessels in 3(8.33%),skin thickening in 4(8.73%).Conclusion The direct and indirect digital molybdenum target mammographic signs of breast cancer are important for early diagnosis and differential diagnosis of breast cancer.

Clinical effect of breast augmentation by transplantation of autologous fat granules / 中华医学美学美容杂志

Objective To investigate the method and clinical effect for transplanting auto-fat granules to augment breast. Methods The autologous fat granules were obtained by liposuetion using syringe from the sites with cumulate fat. After repeated wash and purification, they were transplanted into the interspace behind the breast by injection. The quantity of auto-fat granules injected was 50 - 100 ml in each side of breast per transplantation at 3 - 6 months intervals, and the whole course of treatment needed 2 -6 transplantations. Results 91 of 107 cases were followed up from 6 months to 3 years after final transplantation. All of them were satisfied with the postoperative effect, showing per-fect and plump appearance of the breasts. However, small indurations were found sporadically in 13 cases (21 breasts) within 2 -9 months, and calcifications in 8 cases (11 breasts) within 8 - 17 months after the first transplantation. Conclusion Both patients and surgeons are almost satisfied with the re-sults. The transplantation of autologous fat granules for breast augmentation is an effective and practi-cal method but the postoperative effect is influenced by multiple factors. A few of cases present com-plications after transplantation, so more clinical observation should be taken.

Construction of rat CXCR4 gene lentiviral RNA interference vector and its expression in mesenchymal stem cells / 生物工程学报

To construct the lentiviral RNA interference (RNAi) vector of rat CXCR4 gene, three target sequences were selected according to rat CXCR4 mRNA sequence, the complementary DNA contained both sense and antisense oligonucleotides were designed and synthesized. After phosphorylation and annealing, these double-strand DNA were cloned to Bgl II and Hind III sites of pSUPER. Then the product pRiCXCR4 was confirmed by electrophoresis and sequencing. Next, CXCR4 shRNA was cloned to a transfer vector of lentivirus, pNL-EGFP, and pNL-RiCXCR4-EGFP was produced. It was confirmed by digestion and sequencing that CXCR4 shRNA expression structure was correctly cloned to pSUPER and pNL-EGFP respectively. Three plasmids, pNL-RiCXCR4-EGFP, pHELPER and pVSVG were cotransfected into 293T to package lentivirus particles. The functional titer of obtained virus was determined by flow cytometry after transduction in 293T, the resulting functional titer of unconcentrated virus and concentrated virus were 6.4 x 10(4) TU/mL and 6.9 x 10(6) TU/mL respectively. After the rat mesenchymal stem cells (rMSCs) were transduced with the constructed lentiviral vectors, real-time RT-PCR, Western blotting and flow cytometry were used to evaluate the level of CXCR4 expression. Compared with control group, the CXCR4 mRNA expression were obviously suppressed in all three experimental groups (rMSCs-CXCR4a, rMSCs-CXCR4b, rMSCs-CXCR4c), especially the expression rate in rMSCs-CXCR4b group was reduced by 95.6%. The RNAi lentivirus vector of rat CXCR4 gene has been constructed successfully. This greatly facilitates the further studies such as evaluation the role of CXCR4 in rMSCs recruitment to damaged tissue.

Research in effect of continuing nursing care on patients with rheumatoid arthritis / 中国实用护理杂志

Objective To discuss the effect of continuing nursing care on patients with rheumatoid arthritis. Methods 117 patients with rheumatoid arthritis were divided into the control group (60 pa-tients) and the experimental group(57 patients). The control group adopted clinic follow-up visit and rou-tine responding measures. The experimental group was given knowledge education according to the ASMP course, which included demonstration,nurse-patient communication,communication between patients, video show, telephone follow-up and support, etc. The degree of pain,cognition degree of disease knowledge, fre-quency of physical exercises,compliance with doctor's advice, readmission rate and self-care ability of pa-tients with joint deformity were compared by χ2 test between the two groups. Results The experimental group was superior to the control group in the following aspects, such as degree of pain,cognition degree of disease knowledge, frequency of physical exercises,compliance with doctor's advice, readmission rate and self-care ability. Conclusions Application of continuing nursing care in patients with rheumatoid arthri-tis contribute to the prognosis of patients.

Therapeutic efficacy of lentiviral vector mediated BDNF gene-modified MSCs in cerebral infarction / 生物工程学报

Pretreatment with brain-derived neurotrophic factor (BDNF) reduces ischemic damage after focal cerebral ischemia, and bone marrow mesenchymal stem cells(MSCs) were reported to ameliorate functional deficits after stroke in rats. Here we investigate the synergistically therapeutic effects of BDNF gene-modified MSCs on cerebral infarction. We transfected MSCs with the BDNF gene using a lentivirus-based system and investigated whether the BDNF-modified MSCs contributed to improved functional recovery in a rat transient middle cerebral artery occlusion (MCAO) model. Compared to untreated rats, rats that received both MSCs and BDNF-MSCs showed significantly more functional recovery. The difference in modified neurological severity score(mNSS) was statistically significant (P < 0.001). Recovery was better in BDNF-MSCs than in MSCs (P < 0.001). At the second week and second month after the systemic delivery of blank vector-modified MSCs and BDNF-modified MSCs, the treated rats exhibited more significant recovery than the control, including the accumulation and living of enhanced green fluorescence protein (EGFP)-positive cells in the infarct area and surrounding areas, neuron-like changes, expression of surface markers of neural cells, and a large amount of BDNF expression in the BDNF-MSCs-treated group. Our findings suggest that BDNF-gene-modified rMSCs can migrate to surrounding areas of the cerebral infarction lesion, differentiate into neural cells, and survive for extended periods. With the synergy of BDNF, they may promote the recovery of the neurological function following cerebral infarction and represent a new strategy for stem cell-based therapy.

Effect of SDF-1/CXCR4 on the hematopoietic-supporting activity of bone marrow mesenchymal stem cells / 基础医学与临床

Objective To study the effect of stromal cell-derived factor-1(SDF-1)/CXCR4 on the proliferation of CD34+ hematopoietic stem/progenitor cells(HSPCs) supported by bone marrow mesenchymal stem cells(MSCs).Methods Rat bone marrow MSCs were plated as feeder layer in long-term cultures(LTC) with bone marrow CD34+ cells in vitro.Cultures were weekly supplemented with SDF-1,anti-SDF-1 antibody,or anti-CXCR4 antibody separately for 5 weeks.The hematopoietic-supporting activity was evaluated by the number of CD34+ cells and colony forming cell(CFC).To assess the effect of SDF-1/CXCR4 on CD34+ cell proliferation cycle,killing assays were carried out and proliferation indexes were calculated.Expression of SDF-1 and CXCR4 in MSCs and CD34+ cells were determined by flow cytometry.SDF-1 in MSC-and CD34+ cell-conditioned medium was also measured by ELISA.Results The number of CD34+ cells,CFC and proliferation indexes were significantly increased in treat-ment with SDF-1(P

Construction of lentiviral vector carrying the Ang-1 gene and its expression in the rMSCs / 中国病理生理杂志

AIM: To construct lentiviral vector carrying the angiopoietin-1(Ang-1) gene,and make it express Ang-1 in the rat mesenchymal stem cells(rMSCs).METHODS: The cDNA encoding the CDS of Ang-1 gene was obtained from the placenta of the adult Fisher 344 rats with RT-PCR.After digestion with restrication endonuclease,the Ang-1 gene was recombined to construct the transfer plasmid PNL-Ang1-IRES2-EGFP.The three-plasmid system of lentiviral vector was consisted of PNL-Ang1-IRES2-EGFP,the packaging plasmid HELPER,and the envelope plasmid VSVG,which were co-transfected to 293T cells mediated by lipofectamin2000 to produce lentiviral particles.The rMSCs were infected by obtained lentiviral particles.The insertion of Ang-1 gene was detected by PCR,the mRNA expression of Ang-1 in rMSCs was detected with RT-PCR,the protein expression of Ang-1 was observed with immunocytochemistry and Western blotting methods.RESULTS: The result of sequencing showed that the cloned Ang-1 gene was consistent with the sequence reported in GenBank.After digestion with restrication endonuclease,the 1 512 bp fragment of Ang-1 gene and the 10.5 kb vector fragment of PNL-IRES2-EGFP were observed with gel electrophoresis.The insertion of Ang-1 gene in viral genome was confirmed.The EGFP expression was observed with the fluorescent microscope.In infected rMSCs,the mRNA and protein expressions of Ang-1 were confirmed.CONCLUSION: Lentiviral vector carrying Ang-1 gene has been successfully constructed.The infected rMSCs are able to express the Ang-1 mRNA and Ang-1 abundantly.This will facilitate the following exploratory development of Ang-1 gene-modified rMSCs.

Recent thymic output function and T cells immune reconstitution after transplantation / 肿瘤研究与临床

Successful hematopoietic stem cell transplantation depends on T cells immunol reconstitution. At present, the T-cell receptor excision DNA circles(TRECs)is the reliable index to monitor recent thymic output function. Quantitative analysis of TRECs level can determine the recent thymic output function and evaluate T cells immune reconstitution after transplantation.

Study the relationship between the monocyte chemoattractant protein-1 gene -2518G/A polymorphism and cerebral infarction of Han population in Chinese Hunan district / 临床神经病学杂志

Objective To investigate the relationship between monocyte chemoattractant protein-1 (MCP-1) gene -2518G/A polymorphism and cerebral infarction(CI) of Han population in Chinese Hunan district.Methods The -2518G/A polymorphism in MCP-1 gene was detected by polymerase chain reaction-restriction fragment length polymorphism analysis (PCR-RFLP) and DNA sequencing in 162 patients with CI (CI group) and 150 normal controls (NC group). The serum level of MCP-1 in the two groups was determined by enzyme-linked immunosorbent assay (ELISA).Results In CI group, genotypic frequency of GG was 39.5%, GA was 37.0%, and AA was 23.5%. The allele frequency of G was 58.0% and A was 42.0%. In NC group, genotypic frequency of GG was 27.3%, GA was 39.3%, and AA was 33.4%. The allele frequency of G was 47.0% and A was 53.0%. The frequencies of MCP-1 -2518GG genetype and G allele in CI group were significantly higher than those in CI group (all P