CRISPR/Cas9-mediated targeted gene correction in amyotrophic lateral sclerosis patient iPSCs

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1 and FUS mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis of motor neurons derived from SOD1 and corrected iPSCs revealed 899 aberrant transcripts. Our work may shed light on discovery of early biomarkers and pathways dysregulated in ALS, as well as provide a basis for novel therapeutic strategies to treat ALS.

Modeling xeroderma pigmentosum associated neurological pathologies with patients-derived iPSCs

Xeroderma pigmentosum (XP) is a group of genetic disorders caused by mutations of XP-associated genes, resulting in impairment of DNA repair. XP patients frequently exhibit neurological degeneration, but the underlying mechanism is unknown, in part due to lack of proper disease models. Here, we generated patient-specific induced pluripotent stem cells (iPSCs) harboring mutations in five different XP genes including XPA, XPB, XPC, XPG, and XPV. These iPSCs were further differentiated to neural cells, and their susceptibility to DNA damage stress was investigated. Mutation of XPA in either neural stem cells (NSCs) or neurons resulted in severe DNA damage repair defects, and these neural cells with mutant XPA were hyper-sensitive to DNA damage-induced apoptosis. Thus, XP-mutant neural cells represent valuable tools to clarify the molecular mechanisms of neurological abnormalities in the XP patients.

Significance of urodynamics detection in benign prostatic hyperplasia patients with cerebral vascular accident / 中国医师进修杂志

Objective To explore the significance of urodynamics detection in benign prostatic hyperplasia (BPH) patients with cerebral vascular accident. Methods The results of urodynamics were detected in 60 BPH patients with cerebral vascular accident (research group) and 60 simple BPH patients (control group), and the results were compared. Results The levels of bladder compliance (BC), maximum bladder capacity (MCC), maximum urinary flow rate (Qmax), detrusor pressure before voiding (Pdet), maximal urethral closure pressure (MUCP), functional urethral length (FUL) in research group were significantly lower than those in control group:(25.0 ± 4.4)ml/cmH2O (1 cmH2O=0.098 kPa) vs. (34.0 ± 5.9) ml/cmH2O, (203.0 ± 16.8) ml vs. (256.0 ± 20.3) ml, (6.70 ± 0.25) ml/s vs. (11.10 ± 0.43) ml/s, (81.0 ± 13.6) cmH2O vs. (108.0 ± 20.2) cmH2O, (60.0 ± 9.8) cmH2O vs. (88.0 ± 12.2) cmH2O, (3.0 ± 0.5) cm vs. (5.2 ± 0.8) cm, P < 0.05. The level of postvoid residual volume (PRV) in research group was significantly higher than that in control group:(83.0 ± 9.4) ml vs. (48.0 ± 8.3) ml, P<0.05. The rate of low compliance bladder and unstable bladder in research group were significantly higher than that in control group:63.3%(38/60) vs. 25.0%(15/60), 66.7%(40/60) vs. 28.3%(17/60), P<0.01. Conclusions Cerebral vascular accident can increase the bladder dysfunction of patients with BPH .

Enantiomeric characterization and structure elucidation of Otamixaban / 药物分析学报

Otamixaban is a potent (Ki ? 0.5 nM) fXa inhibitor currently in late-stage clinical develop-ment at Sanofi for the management of acute coronary syndrome. Being unproductive in obtaining a suitable crystal of Otamixaban, the required enantiomeric characterization has been accomplished using vibrational circular dichroism (VCD) spectroscopy. Selected by a spectrum similarity index, the calculated spectra of several higher energy conformers were found to match well with the observed spectra. The characteristic IR bands of these conformers were also identified and attributed to the solvation effect. Combined with both the single crystal x-ray diffraction results for an intermediate and the proton NMR study, the absolute configuration of Otamixaban is unambiguously determined to be (R,R).

Clinical effect of elastics draw off balloon catheter for 24 hours on preventing bleeding after transurethral plasmakinetic resection of prostate / 中国综合临床

Objective To investigate the clinical effect of elastics drawing off balloon catheter for 24 hours on preventing bleeding after transurethral plasmakinetic resection of prostate (TUPKRP).Methods Seven hundred patients with benign prostatic hyperplasia (BPH) underwent TUPKRP in the Eighth People's Hospital of Shanghai from Jan.2005 to Jun.2013 were randomly divided into control group (298 cases) and experiment group (402 cases).Patients in control group after transurethral resection of the bladder were given treatment as follow:the bladder was placed with F22 cavity catheter,which were fixed on the medial femoral traction;prostate wound edge was pressed for 2-3 d and saline continuous irrigated bladder for avoiding infection.Patients in experiment group were given the same treatment with control group beside prostate wound pressed for 24 h.The days (1st,3rd,5th) of bleeding after operation,cases of hematuria turn clear,cases of postoperative blood transfusion and operation bleeding were recorded.Results Cases of hematuria disappeared postoperation at 1 st,3rd,5th days after operation were 52,42,24 cases in control group and 34,22,10 cases in experiment group.The differences were significant(x2 =6.608,9.279,7.624 ;P =0.010,0.002,0.006).Cases of blood transfusion and reoperation in experiment group were 6 and 2 cases,lower than that in control group(15,9 cases ; x2 =4.955,5.264 ; P =0.026,0.022).Conclusion The method of elastics drawing off balloon catheter for 24 hours improved the effect of preventing bleeding after TUPKRP.

Influence of curcumin in NO and S1 00βlevels in brain tissue of rats after focal cerebral ischemia reperfusion inj ury / 吉林大学学报(医学版)

to observe the infarction volume.Nitrate reductase assay was used to detect the level of NO in brain tissue of the rats.The level of S100βin brain was detected by ELISA method.Results Compared with model group,the brain infarction volumes of the rats 24 and 72 h after cerebral ischemia reperfusion in curcumin group were significantly decreased (P<0.05).Compared with sham operation group,the NO and S100βlevels in the brain tissue 24 and 72 h after cerebral ischemia reperfusion of the rats in model group were significantly increased(P<0.05);compared with model group,the levels of NO in the brain tissue 24 and 72 h after cerebral ischemia reperfusion in curcumin group were remarkably decreased (P<0.05);compared with modee group,the level of S100βin the brain tissue 72 h after cerebral iscemia reperfusion in curcumin group was remarkably decreased (P < 0.05 ). Conclusion Curcumin can significantly reduce the degree of ischemia reperfusion injury in the rats and reduce the levels of NO and S100βin brain tissue,which suggests that the decrease of NO and S100βlevels in brain tissue may be associated with the neuroprotective effect of curcumin.

The influence of post-void residual volume on prostate specific antigen in patients with benign prostatic hyperplasia / 中国医师进修杂志

Objective To study the influence of post-void residual volume on prostate specific antigen (PSA) in patients with benign prostatic hyperplasia (BPH).Methods One hundred and sixty BPH patients who diagnosed by postoperative pathologic manifestations were selected.The patients were divided into 4 groups according to post-void residual volume with 40 cases each:non post-void residual volume (post-void residual volume ＜ 60 ml) group,less post-void residual volume (60 ml≤post-void residual volume ＜ 120 ml) group,middle post-void residual volume (120 ml ≤post-void residual volume ＜ 180 ml) group,mass post-void residual volume (post-void residual volume ≥180 ml) group.The serum PSA levels of 4 groups were observed.Results The PSA level in mass post-void residual volume group [(8.52 ± 6.72) iμ g/L] was significantly higher than that in non post-void residual volume group [(5.26 ± 4.16) μ g/L] and less post-void residual volume group [(5.93 ± 5.20) μ g/L],there were statistical differences (P ＜ 0.05),there was no statistical difference between non post-void residual volume group and less post-void residual volume group (P ＞ 0.05).Conclusions The serum PSA level in BPH patients with non post-void residual volume has no change,while the serum PSA level in BPH patients with mass post-void residual volume is significantly elevated.

Global DNA methylation and transcriptional analyses of human ESC-derived cardiomyocytes

With defined culture protocol, human embryonic stem cells (hESCs) are able to generate cardiomyocytes in vitro, therefore providing a great model for human heart development, and holding great potential for cardiac disease therapies. In this study, we successfully generated a highly pure population of human cardiomyocytes (hCMs) (>95% cTnT(+)) from hESC line, which enabled us to identify and characterize an hCM-specific signature, at both the gene expression and DNA methylation levels. Gene functional association network and gene-disease network analyses of these hCM-enriched genes provide new insights into the mechanisms of hCM transcriptional regulation, and stand as an informative and rich resource for investigating cardiac gene functions and disease mechanisms. Moreover, we show that cardiac-structural genes and cardiac-transcription factors have distinct epigenetic mechanisms to regulate their gene expression, providing a better understanding of how the epigenetic machinery coordinates to regulate gene expression in different cell types.

Astragalus injection strengthens biological viability of rat neural stem cells / 中国组织工程研究

BACKGROUND: Neuroscience and brain science researches have paid attention to the effect of astragalus membranaceus in the treatment of neurologic impairment disease and neural regeneration. Studying astragalus membranaceus effects on neural stem cells are becoming a new research direction. OBJECTIVE: To explore the effects of astragalus injection on biological viability of rat neural stem cells. METHODS: Neural stem cells of Wistar rats were separated and cultured. Immunofluorescence staining was applied to identify the neural stem cells. The purified cells were gained by the second subcultivation in vitro, and then the cells were randomly divided into control group and astragalus injection groups with various concentrations (50, 200, 400 g/L) to culture for 6, 12 and 24 hours. The activity of cells was tested by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, and then the immunohistochemistry was applied to detect the expressions of neuron-specific enolase and glial fibril ary acidic protein in the 50 g/L astragalus injection group after induced for 7 days. RESULTS AND CONCLUSION: The viability of neural stem cells increased significantly after intervention with different concentrations of astragalus injection for 6 hours as compared with the control group (P 0.05). Compared with the control group, the cells in the 50 g/L astragalus group differentiated rapidly, and the number of positive cells for neuron-specific enolase was increased significantly (P < 0.05). The neural stem cells proliferation was hastened, and its differentiation was promoted by the interference of astragalus injection.

Expression and clinical significance of a disintegrin and metalloproteinase-12 and proliferating cell nuclear antigen in bladder carcinoma / 中国综合临床

Objective To investigate the expression of a dismtegnn and metalloproteinase-12 (ADAM12) and proliferating cell nuclear antigen (PCNA) in human bladder carcinoma,and to explore their correlation with different grades and stages of bladder cancer.Methods Biopsies of 12 normal bladder and 43 bladder tumors were performed.And immunohistochemistry was conducted to detect the expression of ADAM12 and PCNA in the biopsies.Results Positive expression signals of ADAM12 were detected significantly higher inthe bladder cancer biopsies than that in the normal ones (Z =4.879,P ＜ 0.05),and the expression level of ADAM-12 in lower histological grade was significantly higher than that in the moderate and higher histological grades (x2 =22.3685,P ＜ 0.01).Positive expression signals of PCNA were detected significantly higher in the bladder cancer biopsies than that in the normal ones (Z =4.879,P ＜ 0.05)).Those with lower histological grade had a higher expression level of PCNA when compared with the moderate and higher histological grades (x2 =10.665,P =0.0137).The expression of ADAM-12 was positively correlated with PCNA in bladder cancer (r =1.000,P ＜ 0.0001).Conclusion The over expression of ADAM12 and PCNA maybe play an important role in development of the bladder tumors.And ADAM12 may be a promising biomarker of bladder cancer in the clinical behavior.

Higher-order genomic organization in pluripotent stem cells

Recent applications of new tools for genome-wide mapping of long-range and spatial interactions have shed light onto the fundamental mechanisms of three dimensional chromatin organizations in pluripotent stem cells and their derivatives.

Mini-invasive treatment of cystitis glandularis with affected ureteric meatus / 中国综合临床

Objective To investigate the minimal invasive management of cystitis glandularis with invasion of the ureteric meatus.Methods The clinical data of 18 cases were reviewed.Among the 18 cystitis glandularis patients,12 cases were invasion of bilateral ureteric meatus and 6 of unilateral ureteric meatus.Operation or drug treatment was performed on the diseases that can cause cystitis glandularis such as bladder stone,bladder neck stegnosis,external urethral meatus stegnosis and benign prostate hyperplasia.Sensitive antibiotic was administrated in all cases.After placing ureter catheter,transurethral plasma electro-resection was carried out in five patients whose ureteric meatus could be identified.In addition,of thirteen patients with ureteral orifice unable to be identified,there were ten cases with normal renal function,mitomycin was injected under affected membrana mucosa,and then the patient with ureteral orifice identified underwent transurethral plasma electro-resection after placing ureter catheter.On the other hand,the patient whose ureteral orifice still could not be recognized undertaken transurethral electro-resection at first,during which the ureter catheter was put once ureteral orifice had been detected,otherwise,the ureter catheter should be placed through cystoscope if nephritic colic emerged and hydronephrosis aggravated after operation.In those patients with kidney dysfunction,the ureter catheter was put by ureter discission or ureter replantation at first,the transurethral electro-resection could not be executed until the renal function recovered.Following all these procedure above,bladder instillation of drugs regularly,anti-infection and symptomatic treatment were administrated.Results One patient combined with bladder adenocarcinoma received cystectomy,of the other patients,six cases recurred and underwent electrotomy again resulting in no relapse.All nephrohydrops vanished or relieved obviously,nevertheless,urinary tract infection,haematuria and bladder-ureter backstreaming as the cardinal complication developed in some cases.Conclusion In the management of cystitis glandularis encroaching ureteric meatus,total or partial cystectomy can be avoided if ureter draining freely can be ensured,motivation removal,antiinfection,injection of drug under mucosa and preoperative diuresis conduce to the achievement of ureter catheter placing,transurethral plasma electro-resection is still effective methods in treating these cystitis glandularis.

Donor cytokine gene polymorphisms affect the development of acute rejection after kidney transplantation / 中华器官移植杂志

Objective To investigate the influence of renal allograft donor cytokine and cytokine receptor gene polymorphisms on acute rejection after renal transplantation.Methods (1) 126 cases of cadaveric renal allograft recipients were divided into two groups according to the presence or absence of acute graft rejection.The distribution of 22 polymorphisms in 13 cytokine genes and production types of some cytokines were compared between donors of two groups as well as latent factors affecting acute rejection.(2) Based on the result of HLA-DR matching,all recipients were stratified into two conditions:0～1 locus HLA-DR mismatched and HLA-DR completely mismatched.Previous positive gene polymorphisms were compared between rejection group and no rejection group under two conditions.Results (1) Compared with no rejection group,the number of HLA-DR mismatched was significantly higher in rejection group.In the donors of rejection group,the genotype frequency of IL-1α-889 C/C,IL-1Rα msp I 11100 T/T,IL-4Rα+ 1902 A/A,TGF-β1 codon 10 C/C,IL-10-1082 A/A and lower production type frequency of IL-10 were significantly higher,whereas the genotype frequency of IL-12-1188 A/A,IL-2-330 G/G and IL-10 GCC/ATA was significantly lower.(2) With 0～1 locus HLA-DR mismatched,the genotype frequency of IL-1Rα msp I 11100 T/T,IL-4Raα+1902 A/A,IL-2-330 G/G,TGF-β1 codon 10 C/C and lower production type frequency of IL-10 showed significant difference between two groups,whereas with HLA-DR completely mismatched,the genotype frequency of IL-12-1188 A/A had significant difference.Conclusions This study verifies renal allograft donor genotype of IL-1α-889 C/C,IL-4Rα+ 1902 A/A,IL-1Ra msp I 11100 T/T,TGF-β1 codon 10 C/C,IL-10-1082 A/A and lower production type of IL-10 gene as the genetic safe factors for the development of acute allograft rejection and genotype of IL-12-1188 A/A and IL-2-330G/G as the major genetic risk factors for acute rejection after kidney transplantation Furthermore,the condition of HLA-DR mismatching might interfere with the action of these cytokine and cytokine receptor polymorphisms.

Comparison of the clinical effectiveness of transurethral resection of the prostate and transurethral vaporization-resection of the prostate / 中华泌尿外科杂志

Objective To compare the efficacy and complications between transurethral resec-tion of the prostate(TURP) and transurethral vaporization-resection of the prostate (TUVRP). Methods 637 cases of benign prostatic hyperplasia(BPH)were divided into 2 groups, 298 cases un-derwent TURP and 339 underwent TUVRP. The maximal flow rates (Qmax) were (9.8±2.3)ml/s, (10.1±2.1) ml/s, the international prostatic symptom scores (IPSS) were 15.3±3.1,15.1±3.7 re-spectively. The surgical outcomes and the complications of the 2 groups were analyzed. Results For the TURP group and TUVRP group, the postoperative Qmax were (19.0±2.9)ml/s and (18.0±2.3) ml/s, both significantly higher than those of preoperation(P<0.01). There was no significant differ-ence between the 2 groups(P0.05). For the TURP group, the mean operation time was (52±16) rain, visual hematuria lasting for (9.0±2.3) d, secondary bleeding in 6 cases (2.0%), lower urinary tract infection in 14 cases(4.7%), 1 month after operation lower urinary tract symptom(LUTS) in 26 case(8.7%), IPSS was 5.0±1.4, contemporary incontinence in 6 cases(2.0%), memberanous sticture 4 cases(1.3%). For the TUVRP group, the mean operation time was (68%19)min, visual hematuria lasting for (12.0±3.6) d, secondary bleeding in 19 cases (5.6%), lower urinary tract infection in 38 cases(11.2%) ,1 month after operation LUTS in 59 cases(17.4%) ,IPSS was 8.0±1.6,contemporary incontinence in 13 cases(3.8%), memberanous stieture in 16 cases (4.7%). There were significant differences between the 2 groups. For the TURP group, blood loss during operation was (126±29) ml, resected tissue weighed (31±8)g, pre- and post-operative serum natium consentration balance was (8±6)mmol/L,TURS developed in 3 cases(1%). For the TUVRP group, blood loss during opera-tion was (122±38)ml, resected tissue weighed (33±9)g,pre- and post-operative serum natium con-sentration balance was (7±7) mmol/L, TURS developed in 2 cases(0.6%). There was no significant differences(P0.05). Conclusions TURP and TUVRP have similar efficacy treating symptomatic BPH. The incidences of complications of TUVRP are slightly higher than those of TURP.

Angiogenesis and Neurogenesis Promoted by Vascular Endothelial Growth Factor After Cerebral Ischemia / 国际脑血管病杂志

Vascular endothelial growth factor (VEGF) is a powerful angiogeuetic factor. In recent years, it has been found that it has a powerful capacity to promote nerve regeneration, and has potential value in the treatment of ischemic cerebrovascular disease. Newborn endothelial celts can form "vascular niche", and promote neurogenesis by releasing an array of neurotrophic factors. Also, newborn neuronal cells can enhance angiogenesis. There is a "cross-talk" between angiogenesis and neurogenesis, and VEGF plays a very important intermediary role in it. This article reviews the studies of VEGF in promoting angiogenesis and neurogenesis after cerebral ischemia.

Spontaneous Hemorrhagic Transformation After Cerebral Infarction and Its Imaging Assessment / 国际脑血管病杂志

The research of hemorrhagic transformation (HT) after cerebral infarction is now focus on HT after thrombolytic therapy. However, attention should also be aroused to the occurrence of spontaneous HT in most patients who cannot receive thrombolysis promptly because of various reasons, for it has important influence on the treatment of cerebral infarction and prognosis. This article reviews the concept, typing, pathogenesis, correlation factors, imaging diagnosis and prediction of HT, as well as the effects of non-thrombolytic therapies on HT, such as antiplatelet, defibrase, anticoagulant, anti-free radical, and the relations between HT typing and prognosis.

Fibroblast Growth Factor and Angiogenesis After Stroke / 国际脑血管病杂志

Increasing cerebral blood flow in ischemic penumbra helps to promote neurological function recovery.Fibroblast growth factor is closely associated with angiogenesis after stroke;it may improve cerebral blood flow in ischemia penumbra,and thus contributes to neurological function recovery.Its application will become a novel approach in the treatment of ischemic stroke.

Inhibition of caspase-12 expression by NGF in rabbit cerebral ischemia/reperfusion injury / 基础医学与临床

Objective To observe the effect of NGF on the caspase-12 in cerebral ischemia/reperfusion. MethodsTwenty-six healthy male white rabbits were randomly divided into sham, ischemia/reperfusion and NGF group. The tissues were sampled to detect the expression of caspase-12, caspase-3 by immunohistochemistry, and to detect the cerebral apoptosis by flow cytometry and TUNEL staining methods.Results Compared with sham group, the expression of caspase-12 and caspase-3 in I/R group was higher (P

Theory of the spleen and stomach and AIDS prevention and treatment by TCM / 中华中医药杂志

Theory of the spleen and stomach is an important component of TCM.In the article,the author introduced the principal knowledge of spleen and stomach in TCM,the recognition of AIDS in TCM,the application of theory of spleen and stomach in prevention and treatment for AIDS.It had important significance for theory of spleen and stomach in treating and preventing AIDS.

The diagnosis and treatment of cytomegalovirus (CMV) pneumonia after renal transplantation / 解放军医学杂志

Objective To explore the diagnosis and treatment of cytomegalovirus (CMV) pneumonia in patients who received immunosuppressive therapy after renal transplantation. Methods 15 patients out of the 110 recipients of renal transplantation were diagnosed as to have CMV pneumonia, during a period of the 2nd to 4th month after renal transplantation. All the 15 patients were treated with a systematic strategy, including rational use of antibiotics, withdrawal of immunosuppressant drugs and rebuilding the body′s immunity function, timely application of mechanic ventilator, and continuous high volume hemofiltration (CVVHF). The clinical responses of the 15 patients were reviewed and retrospectively analyzed. Results 15 recipients contracted CMV pneumonia out of 110 cases (13.6%), and it occurred on a mean of 93.3 days after renal transplantation. All of them were febrile with cough, and showed manifestations of interstitial pneumonia on chest films. CMV-IgM were positive in 13 cases, CMV DNA positive in 9 cases, and 4 patients presented signs of acute respiratory distress syndrome. The condition developed quickly in all the patients. 9 patients recovered and 5 patients died, including one who gave up treatment for financial reason. Conclusion CMV infection is a serious complication after renal transplantation. Early diagnosis with detection of CMV DNA and serum antibody of CMV-IgM should be emphasized. The comprehensive protocol principally composed of anti-virus therapy with cymevan or ganciclovir, monitoring of peripheral lymphocyte subgoups, and adjustment of dosage of immunosuppressive agent may improve recovery rate of CMV pneumonia.