The conventional method for the derivation of human embryonic stem cells (hESCs) involves inner cell mass (ICM) co-culture with a feeder layer of inactivated mouse or human embryonic fibroblasts in an in vitro fertilisation culture dish. Growth factors potentially involved in primary derivation of hESCs may be lost or diluted in such a system. We established a microdrop method which maintained feeder cells and efficiently generated hESCs. Embryos were donated for stem cell research after fully informed patient consent. A feeder cell layer was made by incubating inactivated mouse embryonic fibroblasts (MEFs) feeder cells in a 50 μl drop of medium (DMEM/10% foetal calf serum) under mineral oil in a small tissue culture dish. MEFs formed a confluent layer and medium was replaced with human embryonic stem medium supplemented with 10% Plasmanate (Bayer) and incubated overnight. Cryopreserved embryos were thawed and cultured until the blastocyst stage and the zona pellucida removed with pronase (2 mg/ml; Calbiochem). A zona-free intact blastocyst was placed in the feeder microdrop and monitored for ES derivation with medium changed every 2-3 d. Proliferating hESCs were passaged into other feeder drops and standard feeder preparation by manual dissection until a stable cell line was established. Six hESC lines (Shef 3-8) were derived. From a total of 46 blastocysts (early to expanded), five hESC lines were generated (Shef 3-7). Shef 3-6 were generated on MEFs from 25 blastocysts. Shef7 was generated on human foetal gonadal embryonic fibroblasts from a further 21 blastocysts. From our experience, microdrop technique is more efficient than conventional method for derivation of hESCs and it is much easier to monitor early hESC derivation. The microdrop method lends itself to good manufacturing practice derivation of hESCs. 相似文献
In controlled conditions, the effect of leaf infection by Mycosphaerella fragariae on total superoxide dismutase (SOD, EC 1.15.1.1) activity and induction of SOD isozymes was studied in three different strawberry cultivars, i.e. "Joliette" (resistant), "Honeoye" (partially resistant) and "Kent" (susceptible). Infection of the strawberry leaves with M. fragariae resulted in increase in SOD activities in all three cv. Total SOD increased 1d after inoculation in Joliette and Kent, and 2d after inoculation in Honeoye and reached the highest level in all three cv, at the 2nd day after inoculation, then slowly declined afterward. Total SODs in Joliette and Honeoye at the 2nd day after inoculation were 4516 and 4947Ug(-1) FW, respectively, which were significantly higher than that of Kent (3255Ug(-1)FW). Banding pattern of SOD isozymes in all three cv was also affected by infection. Electrophoresis profile of infected cv revealed two newly synthesized isozymes in Joliette and Honeoye, in which one band, i.e. R(f) = 0.53 was observed exclusively in inoculated Joliette and Honeoye. Therefore, it is considered to be associated with leaf spot resistance. 相似文献
Cutaneous Leishmaniasis (CL) is one of the most prevalent clinical forms of leishmaniasis. Preliminary data suggest that cytokine gene polymorphisms can contribute to resistance or susceptibility to CL. Therefore, we investigated the association of functional polymorphisms in four cytokine genes with susceptibility to, and clinical outcome of CL. A total of 201 patients with self-healing cutaneous leishmaniasis (SCL) and 92 asymptomatic infected controls (AIC) from Fars province as well as 58 patients with chronic cutaneous leishmaniasis (CCL) and their 688 normal controls (normal Iranian population or NIP) who were collected from the different areas of Iran were included in the study. The allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) or PCR-RFLP (restriction fragment length polymorphism) methods were used for genotyping. The frequency of TNF-alpha -308 G-->A and TNF-beta +252 G-->A gene polymorphisms were not different between studied groups. Distribution of IFN-gamma +874 A-->T and IL-4 -590 C-->T polymorphism were also compared between SCl or CCL patients and their controls. IFN-gamma +874 A-->T polymorphism was less common in CCL patients compared to the NIP (chi(2)=12.53, p=0.0019). Significant differences in frequency of IL-4 -590 C -->T polymorphism were also found between the SCL and AIC (chi(2)=8.64, p=0.003). In conclusion, our results suggest that functional genetic variants in the IL-4 promoter could influence the risk of developing CL while the polymorphism in the first intron of the IFN-gamma gene might influence the progression of disease towards CCL. 相似文献
Summary The present study diseusses the results of cytological studies of two kinds of sugar beet callus, i.e., embryogenic and non-embryogenic
tissues. The calluses were produced through culture of secondary leaves on Murashige and Skoog medium containing two hormone
combinations. One week after transfer of calluses onto fresh medium, their cells were viewed using electron microscopy and
an image analyzer. Observations showed that cells of the two callus types had considerable differences in cell structure and
various organelles. Of note were the high amount of polyploidization, rough endoplasmic reticulum, polysome, poly-nucleolus,
and incomplete cell wall together with abnormal partitioning in non-embryogenic cells, as compared to embryogenic cells. In
contrast, vacuolation of cytoplasm, perfect cell wall and partitioning structure, and the high proportion of nucleus/cytoplasm
area were recognized in embryogenic cells. 相似文献
Multiple myeloma (MM) is a clonal B‐cell malignancy characterized by the accumulation of neoplastic proliferation of a plasma cell in the bone marrow that produces a monoclonal immunoglobulin. The immune checkpoint inhibitors against programmed death‐1/programmed death‐1 ligand and cytotoxic T‐lymphocyte antigen 4 axis have demonstrated appropriate anticancer activity in several solid tumors and liquid cancers, and are rapidly transforming the practice of medical oncology. However, in a high percentage of patients, the efficacy of immune checkpoints blockade remains limited due to innate or primary resistance. Moreover, the malignancies progress in many patients due to acquired or secondary resistance, even after the clinical response to immune checkpoints' blockade. The evidence shows that multiple tumor‐intrinsic and tumor‐extrinsic factors and alterations in signaling pathways are involved in primary and secondary resistance to immune checkpoints blockade. Improved identification of intrinsic and extrinsic factors and mechanisms of resistance or response to immune checkpoints blockade may not only provide novel prognostic or predictive biomarkers but also guide the optimal combination/sequencing of immune checkpoint blockade therapy in the clinic. Here, we review the underlying biology and role of immune checkpoints blockade in patients with MM. Furthermore, we review the host and tumor‐related factor effects on immune checkpoints blockade in MM immunotherapy. 相似文献
Thyroid cancer is the most common malignant tumor of the endocrine system seen in the thyroid gland. More than 90% of thyroid cancers comprise papillary thyroid cancer (PTC) and follicular thyroid cancer (FTC). Although anaplastic thyroid carcinoma (ATC) accounts for less than 2% of thyroid cancer. But patients’ lifespan after diagnosis is about 6 months. Surgical interventions, radioactive iodine use, and chemotherapy are not sufficient in the treatment of ATC, so alternative therapies are needed.
Methods and results
The WST-1 assay test was performed to evaluate the anti-proliferative effects of Valproic acid (VPA). Also, the effect of VPA on miRNAs affecting histone deacetylase was determined by Quantitative RT-PCR. In the SW1736 cell line, IC50 dose for VPA was found 1.6 mg/ml. In our study, the level of oncogenic genes expression in cells treated with VPA, including miR-184, miR-222-5p, miR-124-3p, and miR-328-3p, decreased. Also, the expression of tumor inhibitory genes including miR-323-5p, miR-182-5p, miR-138-5p, miR-217, miR-15a-5p, miR-29b-3p, miR-324-5p and miR-101-5p increased significantly.
Conclusions
VPA can ad-just countless gene expression patterns, including microRNAs (miRNAs), by targeting histone deacetylase (HDAC). However, further studies are required for more accurate results.
ABSTRACT: Glossopharyngeal neuralgia is an uncommon, painful syndrome, characterized by paroxysmsof pain in the sensory distribution of the 9th cranial nerve. Idiopathic glossopharyngealneuralgia may be due to compression of the glossopharyngeal nerve by adjacent vessels,while secondary glossopharyngeal neuralgia is associated with identifiable lesions affectingthe glossopharyngeal nerve at different levels of its neuroanatomic pathway.Glossopharyngeal neuralgia is rare in the general population, but is more common in patientswith multiple sclerosis. CASE PRESENTATION: A 56-year-old Caucasian woman with multiple sclerosis and migraine presented to ourfacility with intermittent lancinating pain to the right of her throat, tongue, and the floor ofher mouth that had been occurring for the past year. The pain was intense, sharp, andstabbing, which lasted two to six seconds with radiation to the right ear. Initially, the attackswere infrequent, however, they had become more intense and frequent over time. Our patientreported weight loss, headache, painful swallowing, and the inability to maintain sleep due topainful attacks. A neurological examination revealed a right-handed woman with triggerpoints in the back of the tongue and throat on the right side. She also had dysphagia,hoarseness, and pain in the distribution of the right glossopharyngeal nerve. Mild righthemiparesis, hyperreflexia, dysmetria, and an ataxic gait were present. A magnetic resonanceimaging scan of the brain was consistent with multiple sclerosis and magnetic resonanceangiography demonstrated a loop of the posterior inferior cerebellar artery compressing theright glossopharyngeal nerve. She responded satisfactorily to carbamazepine. Microvasculardecompression and Gamma Knife? radiosurgery were discussed in case of failure of themedical treatment; however, she declined these options. CONCLUSIONS: Glossopharyngeal neuralgia in multiple sclerosis may occur due to vascular compressivelesions and it should not be solely attributed to the underlying demyelinating process.Vascular compression of the glossopharyngeal nerve could independently causeglossopharyngeal neuralgia in patients with multiple sclerosis, and vascular imaging toexclude such a diagnosis is recommended. 相似文献
Urea-formaldehyde polymers, which are utilized in the adhesives industry, have recently been shown to be suitable materials for synthesizing micro/nanocapsules for use in self-healing (nano)composites. In this study, molecular dynamics was employed to simulate the process in which urea and formaldehyde are cross-linked via methylene and ether cross linkers, and to study the structure and mechanical/thermal properties of simulated poly(urea-formaldehyde)s (PUFs). The elastic stiffness constants of the simulated materials were calculated using the constant-strain (static) method. A temperature cycle was applied to the cross-linked PUFs, and the glass transition behavior of each material was investigated through the mean squared displacement (MSD) and temperature evolution of the energy and the specific volume of the polymer. The simulation results confirmed that there was considerable improvement in the properties of the poly(UF) materials upon cross linking. The radial distribution function was also used to study the local structures of the polymers, and this revealed that increasing the temperature and cross linking density results in a significant drop in hydrogen bonding intensity in the cross-linked PUF systems. 相似文献