Modeling CADASIL vascular pathologies with patient-derived induced pluripotent stem cells

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a rare hereditary cerebrovascular disease caused by a NOTCH3 mutation. However, the underlying cellular and molecular mechanisms remain unidentified. Here, we generated non-integrative induced pluripotent stem cells (iPSCs) from fibroblasts of a CADASIL patient harboring a heterozygous NOTCH3 mutation (c.3226C>T, p.R1076C). Vascular smooth muscle cells (VSMCs) differentiated from CADASIL-specific iPSCs showed gene expression changes associated with disease phenotypes, including activation of the NOTCH and NF-κB signaling pathway, cytoskeleton disorganization, and excessive cell proliferation. In comparison, these abnormalities were not observed in vascular endothelial cells (VECs) derived from the patient's iPSCs. Importantly, the abnormal upregulation of NF-κB target genes in CADASIL VSMCs was diminished by a NOTCH pathway inhibitor, providing a potential therapeutic strategy for CADASIL. Overall, using this iPSC-based disease model, our study identified clues for studying the pathogenic mechanisms of CADASIL and developing treatment strategies for this disease.

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.

Preliminary Study on Boiling Water Brewing Extraction of Astragalus Polysaccharides / 中国药师

Objective:To study the boiling water brewing extraction of astragalus polysaccharides in order to solve the problems of long cycle , high energy consumption and poor clarity in the traditional extraction , and lay foundation for the industrial production . Methods:Using the extraction yield of astragalus polysaccharide as the evaluation index and phenol -sulfuric acid method applied to de-termine the content of astragalus polysaccharide , the boiling water brewing extraction was used for astragalus polysaccharides .The effects of brewing time , boiling water amount and brewing times on the polysaccharide content were studied by single factor experi -ments, and then an orthogonal design method was used to screen the optimum technology parameters .The traditional water decoction extraction process was employed as the control .Results:The optimum conditions of boiling water brewing were as follows:adding 9-fold amount of water and soaking 60 min for three times.The astragalus polysaccharides extraction yield (4.81) of the optimal technology was higher than that of the traditional water decoction extraction (4.06%).Conclusion:Boiling water brewing method used to extract astragalus is with high extraction yield , simple operation , short cycle and low energy consumption , the color of astragalus polysaccha-ride is light, and it is clear after dissovled in water , which is superior to the traditional water decoction extraction method , and provide a new method with broad application prospect for the preparation of astragalus related preparations .

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.

Ankle-foot orthosis improves walking ability of hemiplegic patients:a Meta-analysis / 中国组织工程研究

BACKGROUND:Studies have shown that ankle-foot orthosis can increase the feedback on the input information from receptors in the skin of the foot and leg to improve the ankle joint position sense, and promote brain function reorganization. OBJECTIVE:To systematical y evaluate the effect of ankle-foot orthosis on the improvement of walking in hemiplegic patients. METHODS:The Chinese Biomedical Literature Database, CNKI, WanFang Data and VIP database were searched for reports of randomized control ed trials of ankle-foot orthosis to improve walking ability in hemiplegic patients, from the date of establishment of each database to June 2013. The randomized control ed trials which met the criteria were included for the Meta-analysis. RESULTS AND CONCLUSION:A total of 9 randomized control ed trials involving 456 patients were included. Meta-analysis showed that, compared with conventional treatment and drug therapy, ankle foot orthosis via the continuous treatment shows certain advantages to improve lower extremity motor function in hemiplegic patients, life skil s and 10-meter maximum walking speed. Due to a limited number of included documents, the remaining indicators such as walking speed, stride difference and balance function were only for appropriate descriptive analysis. The results suggested that, by improving abnormal gait, walking speed, stride frequency, gait cycle, space asymmetry, ankle muscle spasms and balancing, the ankle-foot orthosis could achieve the goal of improving walking function. Ankle-foot orthoses could not be confirmed to exert the role in the fol owing indicators, including time asymmetry, double support phase prolongation and stride length. This evidence shows that ankle-foot orthoses in hemiplegic patients may promote recovery of motor function of the lower limbs and activities of daily living to a certain extent, but the more high-quality, multi-center randomized control ed trials with large samples are necessary.

Effects of Telmisartan on Insulin Resistance and Oxidative Stress in Nonalcoholic Steatohepatitis Rats / 天津医药

Objective To investigate the effects of telmisartan on insulin resistance and oxidative stress in nonalco?holic steatohepatitis (NASH) rats. Methods Fifty male SD rats were randomly divided into five groups:control group, mod?el group, polyene phosphatidylcholine group, low-dose telmisartan group and high-dose telmisartan group by using random number table (n=10 in each group). Control group was given standard food,the other groups were given high fat diet for 12 weeks to establish NASH rat model. Then intervention groups were given either normal saline 1.0 mL/(kg·d) or polyene phos?phatidylcholine 8.4 mg/(kg·d), or telmisartan 4 mg/(kg·d) or telmisartan 8 mg/(kg·d) for 4 weeks by intragastric adminstra?tion. All rats were sacrificed at the end of the 16th week, the lever of plasma insulin resistance index (HOMA-IR), ALT, AST, TG, TC, MDA, SOD, T-AOC, CAT, GSH-PX and liver homogenate MDA, SOD, GSH-PX and liver NAS scores were tested. Results In polyene phosphatidylcholine treated group, the lever of plasma ALT, AST, HOMA-IR and liver NAS scores were degreased significantly compared with model group. The lever of plasma AST, SOD, T-AOC, CAT, GSH-PX and liver homogenate SOD, GSH-PX, liver NAS scores were improved in both low-dose and high-dose telmisartan groups com?pared with model group while plasma and liver homogenate MDA , HOMA-IR were reduced significantly in these two groups compared with model group. Besides, plasma ALT was significantly improved in high-dose telmisartan group compared with model group. Conclusion Telmisartan reduce plasma ALT, AST, oxidative stress, HOMA-IR and liver NAS scores in NASH rats. And high-does telmisartan is better than low-dose telmisartan and polyene phosphatidylcholine in treatment ef?fect.

Sequential therapy of BTD and MPT regimen for the newly-diagnosed multiple myeloma patients no eligible for bone marrow transplantation / 白血病·淋巴瘤

Objective To retrospectively analyze the outcomes and adverse effects of sequential therapy of BTD and MPT regimen for the newly-diagnosed multiple myeloma patients no eligible for high dose chemotherapy and stem cell transplantation. Methods Thirty-six patients were involved in this study and the patients were treated with tandem therapy of BTD and MPT regimen. The patients were treated with BTD regimen as induced therapy no less than 2 cycles. When the patients got PR or above PR,they were treated with MPT regimen as consolidation therapy which was no less than 2 cycles. Then,the patients who achieved PR or partial PR were received MPT chemotherapy regimen as consequent treatment. After that,low dose thalidomide was used as maintenance therapy. The outcomes and adverse effects were retrospectively evaluated. Results Thirty-six patients were treated with BTD regimen as induced therapy. The results were that 7 patients (19.4 %) achieved CR,8 (22.2 %) VGPR,14 (38.9 %) PR and the OR rate was 80.6 %. The patients (n=29) who achieved no less than PR was treated with MPT regimen as consequent therapy. The results were that four patients were in progression and the others were stable. Twenty-five patients were treated with low dose thalidomide as maintenance therapy. The median progression-free survival (PFS) did not reached yet until last follow-up (median follow-up time was 16.5 months). One-year overall survival rate was expected 86.0 % and 3-year expected overall survival rate was 77.0 %. The main regimen-associated toxicities included thrombocytopenia,peripheral neuropathy (PN),Herpes Zoster,gastrointestinal symptoms,anemia,neutropenia,constipation,fatigue,rash and so on. The incidence of grade 3 and 4 adverse events was low. Conclusion Sequential therapy of BTD and MPT regimen can be used as the front-line therapy for the newly-diagnosed multiple myeloma patients no eligible for high dose chemotherapy and stem cell transplantation.

Effectiveness of lenalidomide-based chemotherapy on 25 patients with multiple myeloma / 白血病·淋巴瘤

Objective To evaluate the efficacy and safety of lenalidomide(Len)-based regimens in the treatment to patients with multiple myeloma (MM). Methods Twenty-five multiple myeloma patients who received Len-based regimens were enrolled in this study. There were 5 newly-diagnosed MM patients, 13 relapsed/refractory (rel/ref) MM and 7 MM patients at plateau after treatment. The newly-diagnosed patients received R-PAD regimen, rel/ref MM received R-MD and the patients at plateau(maintenance treatment group) due to more than 2 grade peripheral neuropathy (PN) received Len monotherapy. Results All of the five newly-diagnosed MM achieved complete remission (CR) (100 %) after 2 courses of theatment; of 13 rel/ref patients (76.92 %) achieved overall response, including 3 (23.08 %) achieving CR, 2 (15.38 %) very good partial response (VGPR), 5 (38.46 %) partial response (PR) and the overall response rate was 76.92 %. Two of the other three no response patients had stable disease(SD) and one had progress disease(PO). The seven patients in maintenant treatment group maintained remission in an average follow-up period of 38 weeks and their PN was relieved in various degrees. Conclusion R-PAD regimen is a highly active regimen for newlydiagnosed MM and R-MD regimen is a promising regimen for rel/ref MM, especially for the patients with serious PN; Len may be a good choice as maintenant chemotherapy for the patients with PN.

Study on soluble Fas ligand in the mechanism of immune escape and counterattack of colorectal cancer / 中国免疫学杂志

Objective:To investigate the role of soluble Fas ligand (sFasL) in the mechanism of immune escape and counterattack of colorectal cancer.Methods:ELISA was used to detect the level of sFasL in the sera of colorectal cancer patients and the supernatant of SW480. Transmission electron microscopy (TEM), flow cytometry (FCM) analysis and fluorescence microscopy were used to examine the apoptosis of Jurkat induced by the sera of colorectal cancer patients and the supernatant of SW480. The apoptosis was also studied after pretreatment of Jurkat by Fas blocking antibody ZB4. The supernatant of African green monkey cell line Vero was used as control.Results:The concentration of sFasL in sera of colorectal cancer patients was 12.21?1.14 ?g/L before treatment, the concentration decreased significantly after treatment(P

Ki-67 antigen's expression in benign and malignant neoplasms of colorectum and its clinical significance / 中国癌症杂志

Purpose:To study the expression of Ki-67 antigen in normal mucosae,benign and malignant neoplasms of colorectum and its clinical significance. Methods:33 cases of carcinomas,13 cases of benign neoplasms and 13 cases of normal mucosae of colorectum were collected. Ki-67 antigen was detected using flow cytometry.Results:The Ki-67 antigen positive fraction of normal mucosae,benign and malignant neoplasms of colorectum were 2.65%?1.51%,6.38%?2.51%,18.52%?8.53% respectivly, there being significant difference between the groups(P0.05). Conclusions:Ki-67 antigen expression is a valuable marker to distinguish the normal mucosae,benign and malignant neoplasms of colorectum. Ki-67 antigen expression has no relation with tumor size,type of histopathology,Dukes stage,location,length of illness history,age,sex of the colorectal carcinoma patients.