Effect of Yiqi Jiedu Prescription-containing Serum on Proliferation of mTEC and Regulatory T Cells in Myasthenia Gravis Patients with Thymus Hyperplasia / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Yiqi Jiedu prescription-containing serum on the proliferation of medullary thymic epithelial cells （mTEC） and regulatory T （Treg） cells in myasthenia gravis （MG） patients with thymus hyperplasia. MethodAccording to serological methods，35 SD rats were adaptively fed for one week and randomized into the low-，medium-， and high-dose Yiqi Jiedu prescription groups，control group， and prednisone group，with seven rats in each group， which were then gavaged with the corresponding drugs for one week for preparing the drug-containing serum. The effect of Yiqi Jiedu prescription-containing serum at different concentrations on the proliferation of mTEC and Treg cells were determined by cell counting kit-8 （CCK-8） assay. Besides， the effect of mTEC and Yiqi Jiedu prescription-containing serum on Treg cell proliferation were observed through co-culture. ResultThymocytes were cultured for a period of time. Their mean positive rate revealed by flow cytometry using mTEC characteristic marker Ulex europaeus agglutinin Ⅰ （UEAI） was 92.54%. Treg cells were sorted by magnetic beads. The purity of Treg cells after repeated magnetic bead sorting was as high as 92%. mTEC and Treg cells showed high positive expression rates，and their cell purity met the requirements of subsequent experiments. When the concentration of Yiqi Jiedu prescription-containing serum was 2.5%-15%，it exhibited an inhibitory effect against mTEC and Treg cells. When the concentration was equal to or greater than 20%，it promoted cell proliferation，which was further enhanced with the extension of action time. The results after 48 h of culture showed that compared with the control group，prednisone and low-dose Yiqi Jiedu prescription had no significant effect on the proliferation of these two kinds of cells，but the medium- and high-dose Yiqi Jiedu prescription remarkably reduced their proliferation inhibition rate （P<0.01）. After co-culture with mTEC， the control group was not significantly different from the prednisone group and the low-dose Yiqi Jiedu prescription-containing serum group in the proliferation of Treg cells，while the medium- and high-dose Yiqi Jiedu prescription-containing serum groups significantly lowered the proliferation inhibition rate （P<0.01）. ConclusionYiqi Jiedu prescription-containing serum affects the proliferation of mTEC and Treg cells in MG patients with thymus hyperplasia. Compared with the solely cultured Treg cells isolated from MG patients，the Treg cells co-cultured with mTEC exhibit enhanced proliferation in MG patients，suggesting that mTEC can regulate the proliferation of Treg cells. This effect becomes more obvious after the intervention with Yiqi Jiedu prescription-containing serum，indicating that intervention effect of Yiqi Jiedu prescription on Treg cells can be produced during its treatment of mTEC， which may be one of the mechanisms of Yiqi Jiedu prescription-containing serum in alleviating MG.

Role Of Panchakarma In Myasthenia Gravis– A Case Study

Myasthenia Gravis (MG) is a long-term neuromuscular disease that leads to varying degrees of skeletal muscle weakness. The underlying defect is a decrease in the number of available acetylcholine receptors (AChRs) at neuromuscular junctions due to an antibody mediated autoimmune attack. The most commonly affected muscles are those of the eyes, face and swallowing. The cause of this disease can be understood in view of Upahata Dhatu Ushma, Srotas and Marutha respectively. The present observation was conducted with an objective to find out the efficacy of Ayurvedic management in Myasthenia gravis. Here is a case of 50years old Hindu female who was diagnosed as myasthenia gravis (MG) reported with complaining of slurred speech, low pitch of voice, difficulty to open the mouth and to swallow food and weakness in the both upper limbs since 4months was registered in OPD of SKAMCH&RC, Bengaluru. Considering the signs and symptoms patient was treated on the line of Ardita Vata chikitsa, Sarvanga Abhyanga with Moorchita tila taila, Nasya with Yashtimadhu taila, Gandusha with Erimedadi taila, Jihwa nirlekhana with Vacha churna were done. Treatment shows significant improvement in the symptoms without any side effects.

Myasthenia Gravis.

Myasthenia Gravis (MG) is a relatively rare neurological disease that is associated with the loss of the acetylcholine receptors that initiate muscle contraction. This results in muscle weakness, which can be life threatening. MG results from antibody-mediated, T cell-dependent immunologic attack on the end-plate region of the postsynaptic membrane. Management must be individualized, and may include symptomatic treatment with cholinesterase inhibitors and immune modulation with corticosteroids, azathioprine, cyclosporine, and mycophenolate mofetil. Rapid, temporary improvement may be achieved for myasthenic crises and exacerbations with plasma exchange (PEX) or intravenous immunoglobulin (IVIg). Prognosis is good due to improved diagnostic testing, immunotherapy, and intensive care.

Construction of MuSK-mCherry fusion fluorescent protein and used for detection of MuSK antibodies in myasthenia gravis / 中国免疫学杂志

Objective:To construct a fusion protein of extracellular domain peptide fragment of muscle specific kinase ( MuSK) and fluorescent protein mCherry ,and used as antigen in the detection of antibodies against MuSK ( MuSKAb ) in the sera of patients with myasthenia gravis ( MG).Methods:The mCherry gene was amplified by PCR from vector pRSET-B and cloned into pGEM-T Easy Vector,and furthermore, cloned into Eukaryotic expression vector pMT /BiP/V5-His ( MuSK), which contains MuSK extracellular domain 22-452 amino acid peptide fragment gene to construct the fluorescent fusion protein gene MuSK -mCherry.The recombinant vector was subsequently transfected into drosophila S 2 cells for expression.The expressed fusion proteins were verified in confocal mi-croscope ,and used as antigen in the detection of MuSKAb in sera of MG patents in fluorescence immunoprecipitation test .Results:The fluorescent fusion protein MuSK-mCherry was successfully constructed and expressed.The MuSKAb in sera of patents with MG could be detected in fluorescence immunoprecipitation test using the constructed MuSK-mCherry fusion protein as antigen.Conclusion: It is available to use the constructed fluorescent fusion protein MuSK-mCherry as antigen in fluorescence immunoprecipitation test for the detection of MuSKAb in sera of patents with MG.

Association of gene polymorphisms of rapsyn exons with myasthenia gravis / 中国免疫学杂志

Objective:To investigate the association of single nucleotide polymorphisms (SNPs) of receptor-associated protein at the synapse ( rapsyn ) with myasthenia gravis ( MG ).Methods: The genomic DNA was extracted from peripheral blood cells , sampled from 132 patients with MG and 153 control individuals.The 8 exons of rapsyn gene were amplified by PCR ,then the products of PCR sequenced directly.Each sequence was compared with wild-type rapsyn gene , and the association between mutation and clinical symptoms of MG analysed.Results:No mutation was found in the exons 1,2,4,5,6,7,and 8 of rapsyn gene both in MG patients and control group compared with the wild-type rapsyn gene.However,a new SNP,L222R[CTG>CGG(2)] or T665G,was found in exon-3.The allele and genotype frequencies of SNP L 222R met Hardy-Weinberg genetic equilibrium (P>0.05),indicating the group repre-sentativeness.The allele frequencies of G were not statistically different between patient and control groups ( P>0.05 ).There were differences in the 3 genotypes TT , TG and GG between patient ( 42.4% vs 48.5% vs 9.1%) and control ( 49.0% vs 33.3% vs 17.6%) groups ( P<0.05 ).The genotype frequencies of GG were statistically higher in control group than that in patient group , showing a recessive model of inheritance.Conclusion: The SNPs in the rapsyn gene are associated with MG in this study.L222R ( T665 G) is a new SNP found and allele G might be a protective factor for MG.

Thymoma Associated with Myasthenia Gravis in Infancy.

Thymomas are associated with various autoimmune disorders in adults, most commonly with myasthenia gravis (MG). In MG antibodies developed against acetylcholine (ACh) receptors, resulting weakness of muscle after repeated use and recovery following a period of rest. However, MG due to thymoma is extremely rare in pediatric age group.1We report an infant with MG due to invasive thymoma.

The correlation between interleukin-4,15 as well as 18 and telomerase activity in myasthenia gravis / 中国免疫学杂志

Objective:To detect the serum levels of IL-4,IL-15 and IL-18 in myasthenia gravis( MG) and investigate the correlations between telomerase activity of CD4~+ T lymphocytes and these interleukins.Methods:The serum levels of IL-4,IL-15 and IL-18 were detected with ELISA and telomerase activity was detected by polymerase chain reaction enzyme-linked immonosorbent assay(PCR-ELISA) in 30 MG patients and 30 negative controls.Results:The levels of IL-4,IL-15,IL-18 and telomerase of CD4~+ T lymphocytes in MG were significantly higher than those in negative controls (P<0.01).There were positive correlations between the serum levels of IL-4,IL-15,IL-18 and telomerase activity of CD4~+ T cell in MG patients (P<0.01).Conclusion:IL-4,IL-15,IL-18 and telomerase activity of CD4~+ T lymphocytes are involved in the pathogenesis of MG;IL-4,IL-15 and IL-18 may upregulate the telomerase activity of CD4~+ T cells.

Discussion on anesthesia of thymectomy for myasthenia gravis / 重庆医科大学学报

Objective:To investigate the anesthetic management of thymectomy for the patients with myasthenia gravis.Methods:Sixty-two patients with myasthenia gravis who underwent thymectomy in our hospital from 1984 to 2003 were retrospectively studied.Of these patients,21 cases received peroral endotracheal intubation and 41 cases pernasal intubation.Intravenous-inhalation combined anesthesia was taken during operation.All of 62 patients with endobronchial tube were transferred to ICU and received mechanical ventilation.Results:Twenty-one patients with perora endotracheal intubation were extubated at 4h-6h after operation.Of these 21 patients,five cases had to be intubated again because of repeated occurrence of myasthenia crisis.Forty-one patients with pernasal intubation did not extubated until mechanical ventilation for 6 hrs to 5 days,and none needed for intubation again,pernasal intubation cases were significantly different with peroral endotracheal intubation cases.The statistics have no difference betwen the trachea incisioned and the dead cases.Conclusion:In anesthetic management of thymectomy for the patients with myasthenia gravis,pernasal endotracheal intubation is convenient for prolonged mechanical ventilation,it can effectively prevent the occurrence of postoperative myasthenia crisis.The perioperative safety precautions,intravenous-inhalation combined anesthesia during operation and strengthening management of respriratory tract are very important.

Clinical study of surgery for patients with myasthenia gravis and hyperthyroidism / 吉林大学学报(医学版)

Objective: To study the clinical therapeutic characteristics of myasthenia gravis (MG) with hyperthyroidism and the effects of surgical procedures for the patients. Methods:Subtotal thyroidectomy,thymectomy,and simultaneous subtotal thyroidectomy and thymectomywere performed on eleven patients with MG and hyperthyroidism. These patients were followedup after the operation. Results :The neck incisions were infected in two of seven type Ⅱ b patientsdue to trachestomy and the infected incisions were surgically healed. The MG symptoms of threecases (1 cases in type Ⅱ. and 2 cases in type Ⅱ b) relapsed in 3 to 6 months after subtotal thy-roidectomy. The MG symptoms of 2 cases (1 case in type Ⅱa and 1 case in type Ⅱ b) relapsed in 8to 12 months after thymectomy. Among six patients treated by simultaneous subtotal thyroidecto-my and thymectomy,the MG symptoms relapsed in a type I case 3 months after the operation,remitted in three case (1 case in type Ⅱ, and 2 cases in type Ⅱb) and improved in two type Ⅱ bcases since the operations. Conclusion:The simultaneous subtotal thymectomy and thyroidectomyfor patients with MG and hyperthyroidism might have better effects,although infective opportuni-ty of the neck incisions increased owing to trachestomy. However,the prognostic effects were notvery satisfactory whether simple subtotal thyroidectomy or thymectomy for the patients with MGand hyperthyroidism was performed.