An 85-year-old woman with Miller Fisher syndrome.

Miller Fisher's syndrome (MFS) commonly presents in the fourth and fifth decades and are rare in people over 70 years. An 85-year-old female with no significant medical history presented with upper extremity anesthesia, ptosis, and unsteady gait. The patient had a history of hypertension and diabetes mellitus. Physical examination showed bilateral total external ophthalmoplegia, areflexia, and cerebellar ataxia. Radiological and laboratory studies were unremarkable. Lumbar puncture showed albuminocytological dissociation. The combined history, physical examination, and lumbar puncture results established a presumptive diagnosis of MFS. Intravenous immunoglobulin was given for 5 days. The patient gradually improved 10 days after the onset of symptoms. Ophthalmoplegia had fully recovered after 6 months. To the best of our knowledge, this case represented the oldest patient with MFS.

Prevalence, distribution, accumulation, and risk of environmental corticosteroids and estrogens in biofilms from the Pearl River Delta.

Biofilms play a significant role in the biogeochemical processing of organic matter and the environmental fate of emerging pollutants. In this study, we investigated the occurrence and distribution of 32 endocrine-disrupting chemicals (EDCs), including 24 environmental corticosteroids (ECs) and 8 environmental estrogens (EEs), in natural biofilms from the Pearl River system. Their association between biofilms and water and environmental risk were assessed. The ECs and EEs ubiquitously occurred in the biofilms, ranging from <0.61-6.57 ng/g and <0.8-2535 ng/g, respectively. Temporally, there was no obvious variance in either ECs or EEs in the biofilms during the winter and summer, and their concentrations exhibited a spatial trend of upward to midstream, descending downstream, and then seaward attenuation at the estuary. For ECs and EEs, the similar levels of field-derived bioconcentration factors (BCFs) (logarithm values: 2.42-2.86 and 2.72-2.98, respectively) and biofilm organic carbon-normalized partitioning coefficients (Kboc) (3.39-3.69 and 3.35-3.95) suggest the comparable potential of accumulation and sorption by biofilms between these two classes of EDCs. In addition, higher values of BCF and Kboc for the EEs were found in winter and were correspondingly comparable to their distribution coefficients (Kd) and Koc derived from suspended particles and sediment, revealing that biofilms are a competitive environmental compartment for capturing EDCs, particularly during the mature period. A positive logKboc-logKow relationship suggests hydrophobic partitioning as a primary interaction mechanism between the biofilm and EEs. Moreover, high risks from biofilm-associated ECs and EEs might have posed to the fluvial ecosystem. This study provides original insights into the occurrence, fate, and risk of ECs in natural biofilms for the first time and demonstrates that biofilms may not only serve as reservoirs but also serve as sentinels for fluvial EDC contamination. These results contribute to the further understanding of the behavior and fate of EDCs in aquatic environments.

Anti-N-methyl-D-aspartate receptor encephalitis with serum anti-thyroid antibodies and IgM antibodies against Epstein-Barr virus viral capsid antigen: a case report and one year follow-up.

BACKGROUND: Anti-N-methyl-D-aspartate receptor encephalitis is an increasingly common autoimmune disorder mediated by antibodies to certain subunit of the N-methyl-D-aspartate receptor. Recent literatures have described anti-thyroid and infectious serology in this encephalitis but without follow-up. CASE PRESENTATION: A 17-year-old Chinese female patient presented with psychiatric symptoms, memory deficits, behavioral problems and seizures. She then progressed through unresponsiveness, dyskinesias, autonomic instability and central hypoventilation during treatment. Her conventional blood work on admission showed high titers of IgG antibodies to thyroglobulin, thyroid peroxidase and IgM antibodies to Epstein-Barr virus viral capsid antigen. An immature ovarian teratoma was found and removal of the tumor resulted in a full recovery. The final diagnosis of anti-N-methyl-D-aspartate receptor encephalitis was made by the identification of anti-N-methyl-D-aspartate receptor antibodies in her cerebral spinal fluid. Pathology studies of the teratoma revealed N-methyl-D-aspartate receptor subunit 1 positive ectopic immature nervous tissue and Epstein-Barr virus latent infection. She was discharged with symptoms free, but titers of anti-thyroid peroxidase and anti-thyroglobulin antibodies remained elevated. One year after discharge, her serum remained positive for anti-thyroid peroxidase and anti-N-methyl-D-aspartate receptor antibodies, but negative for anti-thyroglobulin antibodies and IgM against Epstein-Barr virus viral capsid antigen. CONCLUSIONS: Persistent high titers of anti-thyroid peroxidase antibodies from admission to discharge and until one year later in this patient may suggest a propensity to autoimmunity in anti- N-methyl-D-aspartate receptor encephalitis and support the idea that neuronal and thyroid autoimmunities represent a pathogenic spectrum. Enduring anti-N-methyl-D-aspartate receptor antibodies from admission to one year follow-up but seroreversion of Epstein-Barr virus viral capsid antigen IgM may raise the important issue of elucidating the triggers and boosters of anti- N-methyl-D-aspartate receptor encephalitis.

Up-regulation of TGF-ß1 mRNA expression in peripheral blood mononuclear cells of patients with chronic fatigue syndrome.

BACKGROUND/PURPOSE: It has been shown that the abnormality in immune cells in chronic fatigue syndrome (CFS) patients is closely associated with the participation of TGF-ß. In order to study the relationship between TGF-ß1 and CFS, we investigated the mRNA levels of TGF-ß1 in peripheral blood mononuclear cells (PBMCs) in patients with CFS. METHODS: Fluorescent quantitative real time reverse-transcription polymerase chain reaction (FQ-RT-PCR) was performed to test TGF-ß1 mRNA expression in PBMCs in 63 cases of CFS, 50 cases of disease controls, and 50 cases of healthy controls. RESULTS: The mean value of TGF-ß1 mRNA expression in CFS patients was ΔΔCt=1.17±0.58, which was significantly higher than the disease controls (ΔΔCt=0.07±1.08, df=111, p < 0.01) and the healthy controls (ΔΔCt=0.00±1.63, df=111, p < 0.01). No significant difference was detected between disease and healthy controls (p > 0.05). CONCLUSION: The expression of TGF-ß1 in PBMCs is significantly elevated in patients with CFS. It might be correlated to the pathogenesis of the disease.

The dose-effect of icariin on the proliferation and osteogenic differentiation of human bone mesenchymal stem cells.

Icariin had been reported as a potential agent for osteogenesis, but the dose-effect relationship needed further research to realize the clinical application of icariin. We isolated and purified human bone mesenchymal stem cells (hBMSCs) and stimulated them with different concentrations of icariin. The cytotoxicity of icariin was evaluated by the methylthiazolytetrazolium (MTT) assay method. The proliferation and osteogenic differentiation of such hBMSCs were investigated for different concentrations of icariin. We found that icariin had a dose-dependent effect on the proliferation and osteogenic differentiation of hBMSCs in a suitable concentration range from 10(-9) M to 10(-6) M, but at concentrations above 10(-5) M, the cytotoxicity limited its use. The extremely low cost of icariin and its high abundance make it appealing for bone regeneration.

[Expression of tau-related protein in spinal cord of patients with Alzheimer's disease].

OBJECTIVE: To study the expression of tau-related protein in spinal cord of Chinese patients with Alzheimer's disease. METHODS: Gallays-Braak stain and immunohistochemical study for tau protein (AT8) were carried out in the spinal cord tissue (T2, T8, T10, L2 and S2 segments) of 3 Chinese patients with Alzheimer's disease. Seven age-matched cases without evidence of dementia or neurologic disease were used as controls. RESULTS: Neurofibrillary tangles were identified in the neurons of anterior horn in 2 Alzheimer's disease cases but none was observed in the controls. Tau-positive axons and astroglia were detected in all Alzheimer's disease cases. Tau immunoreactivity in spinal cord of the patients correlated with that in brain tissue. CONCLUSION: The expression of tau-related protein is demonstrated in the spinal cord of Alzheimer's disease patients suggesting that axonal transport defect may play a role in the pathogenesis of Alzheimer's disease.

Effects of the transcription factor Olig1 on the differentiation and remyelination of oligodendrocyte precursor cells after focal cerebral ischemia in rats.

The differentiation and maturation of oligodendrocyte precursor cells (OPCs) is important for remyelination in the central nervous system. Nevertheless, this process is often limited and incomplete in ischemic injury. Oligodendrocyte transcription factor 1 (Olig1) is important for the maturation of OPCs and the repair of demyelinated lesions. However, how Olig1 modulates the development of OPCs or the remyelination associated with ischemic injury remains unclear. The present study aimed to examine alterations in OPCs, and the expression of myelin and Olig1, at different time-points after focal cerebral ischemia using immunohistochemistry and western blot techniques to elucidate the role of Olig1 in the maturation of OPCs and remyelination. The present results showed that the expression of Olig1 significantly decreased at 1 day after middle cerebral artery occlusion (MCAO) and returned to normal levels from day 3 to 28. Additionally, Olig1 was found to translocate into the nucleus following ischemia in the brain. The number of OPCs in the ischemic striatum significantly declined at days 1 and 3 following MCAO, and increased at days 7, 14 and 28 compared with the control. The expression of myelin basic protein, a marker of mature oligodendrocytes and myelin, gradually decreased from day 1 to 7 after ischemia and recovered at day 14 and 28; however, the levels were lower than those in the control group. The present results indicated that the restored normal level of Olig1 following ischemia may play an important role in the maturation of OPCs through its translocation into the nucleus, where it may promote the growth and development of myelin under pathological conditions. However, this endogenous recovery mechanism fails to fully repair the demyelinated lesion. The data of the present study may help clinicians understand the expression pattern of Olig1 and its potential role in endogenous remyelination after ischemia, which may have implications for the treatment of diseases that lead to demyelination.

A patient with Creutzfeldt-Jakob disease with an insertion of 7 octa-repeats in the PRNP gene: molecular characteristics and clinical features.

BACKGROUND: We evaluated the features of neuropathology, abnormal prion protein (PrP) molecules, and clinical data of a Chinese woman diagnosed with familiar Creutzfeldt-Jakob disease (CJD), having 7 octa-repeats inserted with codon 129 methionine homozygote in the PRNP gene. METHODS: Neuropathologic characteristics of the brain were analyzed by hemotoxylin-eosin stain and electronic microscopy. The presence of abnormal PrP in brains was detected by proteinase K and PrP molecules were evaluated by deglycosylation assay. RESULTS: Spongiform degeneration, with diffuse neuronal loss and mild astrocytic gliosis, as well as with profound degeneration of neurons and astrocytes was observed. Proteinase K-resistant PrP was deposited widely in various regions of the brain. Calculation of the glycosylation ratios of proteinase K-resistant PrP molecules identified that the monoglycosyl isomer was predominant. PrP deglycosylation tests allowed for the identification of a predominant 19-kDa PrP signal that represents a partially proteolytic C-terminal segment, a 27-kDa band that represents the full-length wild-type PrP molecule, and a 30-kDa band that probably corresponds to the full-length mutant PrP molecule. CONCLUSION: : Sporadic CJD-like neuropathologic changes and deposits of proteinase K-resistant PrP have been identified in this familiar CJD case with a 168 base pair nucleotide insertion. The clinical features differ from previously reported cases that had 7 octa-repeat insertion, but bear similarities to sporadic CJD.

Detection of human cytomegalovirus in atherosclerotic carotid arteries in humans.

BACKGROUND/PURPOSE: Atherosclerosis plays an important role in stroke. A microbiological infection has been suggested to be involved in the pathogenesis of atherosclerosis, in particular the human cytomegalovirus (HCMV). The objective of this study was to determine the association between HCMV infection and atherosclerosis of the internal carotid arteries in patients of Chinese Han ethnicity with ischemic stroke. METHODS: HCMV DNA and antigen were detected in atherosclerotic internal carotid arteries from 35 patients with ischemic stroke and 20 controls from a Chinese Han ethnic population. Immunohistochemistry, in situ hybridization and polymerase chain reaction were used to detect the HCMV immediate early (IE) and late (L) antigen as well as DNA in the vascular walls. RESULTS: We observed that the proportion of cases that tested positive for HCMV IE but not L antigen and DNA was statistically greater in stroke patients compared with the control population. CONCLUSION: HCMV IE antigen and DNA were associated with the pathological process of atherosclerosis. The vessel wall might be the infection site of the dormant virus.

[Research progress on pathogenesis of temporary osteoporosis in hip].

Transient osteoporosis of the hip(TOH) is classified as a type of bone marrow edema syndrome. TOH is lack of previous study and there is still controversy about his pathogenesis. In recent years, with the development of multi-discipline, such as imaging, pathology, molecular biology, the study has found that the pathological mechanism is complex, while its mechanism is still not clear, which need further research. This paper summarizes the research progress on the pathogenesis of TOH from neurogenic, osteonecrosis, abnormal vascular function, subchondral fracture, heredity and regional acceleration and son on.

Treatment of scrapie pathogen 263K with tetracycline partially abolishes protease-resistant activity in vitro and reduces infectivity in vivo.

OBJECTIVE: To study the possible effect of tetracycline on protease-resistant activity in vitro and infectivity in vivo of a scrapie strain 263K. METHODS: Scrapie pathogens were incubated with tetracycline at different concentrations for various periods of time and protease-resistant PrP signals were evaluated with proteinase K-treatment and Western blots. The preparations treated with tetracycline were intracerebrally inoculated into golden hamsters and typical TSE manifestations were noted. PrPSc in brain tissues of the infected animals was detected by PrP specific Western blot assays. RESULTS: Protease-resistant PrP was significantly reduced in or removed from the preparations treated with tetracycline in a dose-dependant manner. Compared with the control group after incubated for 53.75 +/- 0.50 days, the preparations treated with 5 mmol/L and 20 mmol/L tetracycline prolonged the incubation time of 61.5 +/- 1.73 and 59.5 +/- 0.58 days (P < 0.05). CONCLUSION: Treatment of scrapie pathogen 263K with tetracycline reduces or removes its protease-resistant activity in vitro.

[Advances in the study of bioactivity and structure-function relationship of regulatory peptide AcSDKP].

As a negative regulator of hematopoiesis, AcSDKP is well known to inhibit the proliferation of hematopoietic stem cells and reported to has a biological function in non-hematopoietic cells recently. Its biological activities and structure-function relationship are reviewed in this paper.

[The clinical value of measuring plasma level of very long chain fatty acids in Addison disease].

OBJECTIVE: To determine the level of very long chain fatty acids (VLCFA) in plasma to find out X-linked adrenoleukodystrophy (X-ALD) in patients with Addison disease. METHOD: By using gas chromatography measurement of plasma levels of C(26:0), ratios of C(26:0)/C(22:0) and C(24:0)/C(22:0) was carried out in 36 patients with Addison disease. RESULT: Among the 36 cases, 6 had elevated plasma VLCFA levels; thus the presence of X-ALD was confirmed. CONCLUSION: Misdiagnosis of X-ALD can be reduced by measuring plasma level of VLCFA early in male patients with Addison disease, especially in young ones.

Recent progress in the improvement of the coupling efficiency of "difficult sequences" in peptide synthesis.

Even though chemistry is now in place that potentially allows high coupling efficiencies to be attained, successful coupling is usually a challenge when so-called "difficult sequences" is encountered in peptide synthesis. Some factors that affect the coupling efficiency have been discussed and related methods to overcome those obstacles have been introduced in present review. All suggestions proposed here are valuable and also feasible to improve the coupling completeness in both liquid-phase or solid-phase

[Clinical observation of proximal femoral anti-rotation nail for the treatment of femoral intertrochanteric fracture].

OBJECTIVE: To explore the curative effect and the recessive loss of blooding of PFNA for the treatment of intertrochanteric fractures of femur. METHODS: From January 2012 to January 2015, a total of 49 patients with intertrochanteric fractures of femur were treated with proximal femoral anti-rotation nail including 41 males and 8 females with an average age of 79 years old ranging from 65 to 91 years old. According to the modified Evans type, 1 case was type I, 12 cases were type II, 36 cases were type III. All cases were fresh fractures. Patients had hip pain, movement limited, joint swelling, bruising, extorsion deformity, X-ray and CT examination showed completely fractures. All patients were treated by closed reduction and PFNA internal fixation. Three comminuted fractures using closed reduction were not satisfied, then were treated by limited PFNA fixation after open reduction. RESULTS: The patients' incision got the grade A healing, no complications such as infection and internal fixation failure happened. All patients were followed up from 6 to 36 months (means 22 months). The pain VAS score decreased from preoperative 7.70±1.97 to 1.00±0.26 at 6 months after operation(P<0.01). Harris hip score improved from preoperative 8.70±4.19 to 91.70±5.31 at 6 months after operation(P<0.01). The outcome at 6 months after operation was excellent in 34 cases, good in 7, poor in 1. The fracture healing time was from 8 to 16 weeks with an average of 12 weeks after operation. One patient with osteoporosis, crushing broken, poor compliance, associated with schizophrenia at the same time, appeared with the displacement of the femoral greater trochanter, with conservative treatment for healing. CONCLUSIONS: Intertrochanteric fractures of femur are common in the elderly trauma, in pain relief, recovery of hip function, to provide quality of life for the patients, PFNA achieved satisfactory effect, but its existence is worth to pay close attention to the recessive loss of blood.

[Effects of OGP(10-14) and its derivative G38I on proliferation and differentiation of rat osteoblasts in vitro].

OBJECTIVE: To investigate the effects of the carboxyl end of osteogenic growth peptide (OGP)-OGP((10-14)) and its derivative G38I on the proliferation and differentiation of osteoblasts (OBs). METHODS: Osteoblasts were isolated from the calvariae of newborn SD rats and cultured to G3. OGP((10-14)) or G38I of the concentrations of 10(-15) to 10(-7) mol/L were added to culture medium for 48 hours respectively. The number of cells was counted and MTT analysis was used to examine the proliferation of the cells. The ultrastructure of cells was investigated by electron microscopy. The osteoblasts of G3 were divided into experimental groups, treated with OGP((10-14)) or G38I of the concentration of 10(-11) mol/L for 48 hours, and control group. The alkaline phosphatase activity in the culture medium was measured. The protein expression level of type-I collagen was evaluated by immunohistochemistry. The core binding factor 1 (Cbfa1) and type-I collagen mRNA level of osteoblasts were determined by RT-PCR. RESULTS: With a biphasic effect on, OGP((10-14)) and G38I stimulated the number enhancement of OBs dose-dependently at low concentration and inhibited it at high concentration. The numbers of OB were the highest (37 x 10(4)/ml +/- 7 x 10(4)/ml and 30 x 10(4)/ml +/- 5 x 10(4)/ml respectively) when treated by OGP((10-14)) or G38I of the concentration of 10(-11)mol/L. The rough endoplasm net was flourishing and the secreting vesicle was abounding in the experimental cells. There was calcium crystal in the control cells. The activity of alkaline phosphatase in the culture medium of the OGP (10(-14)) and G38I groups were higher than that in the control group (4.47 U/g and 3.82 U/g vs 2.21 U/g). The protein expression level of type-I collagen was higher and the mRNA levels of Cbfa1 and type-I collagen were higher in the OGP((10-14)) and G38I groups were increased in the experimental groups in comparison with the control group (P < 0.05, P < 0.01, and P < 0.05). CONCLUSION: They stimulated cell number enhancement dose dependently at low concentration and followed by inhibition at high concentration. Just as the native OGP, OGP((10-14)) and its derivative G38I stimulate the proliferation of osteoblasts, and improve their activity, up-regulate the Cbfa1 and type-I collagen mRNA expression levels and increase the collagen synthesis, thus promoting the differentiation and osteogenic effect of osteoblasts.

[Clinical characteristics of patients with chronic fatigue syndrome: analysis of 82 cases].

OBJECTIVE: To analyze the clinical characteristics of Chinese patients suffering from chronic fatigue syndrome (CFS) and provide clinical and laboratory evidence for the study of its etiology and treatment. METHODS: 82 patients with CFS diagnosed based on the CDC criteria 1994 were recruited. History was collected, and physical examination was made. SCL-90 and memory test were used, and Hamilton Anxiety Rating Scale was used to those showing depression and/or anxiety. Laboratory examination, including examination of electrolytes, blood sugar, creatinine, bilirubin, alkaline phosphatase, alanine trasaminase, etc, was conducted. Western blotting was used to detect the protein-24 of Borna disease virus (BDV) in the plasma of 61 patients and 73 healthy controls. High-pressure chromatography was conducted to detect n-6 fatty acids on the membrane of erythrocytes of 42 patients and 37 healthy controls. Plasma L-carnitine in 61 patients and 73 healthy controls was detected by zymological analysis. In different examinations sex and age-matched controls were used. RESULTS: Most of the patients were 21 approximately 50 years old (74/82, 90.24%). No gender difference was found. The patients usually had 4 approximately 6 symptoms besides distinctive fatigue. Descent of remembrance and/or attention was the most conspicuous accompanying symptoms (69/82, 84.15%). Abnormalities in SCL-90 scores were present in 57 patients (69.51%), e.g, somatization existed most commonly (32/82, 39.02%), and anxiety and depression were 20.73% (17/82) and 18.29% (15/82) respectively. The prevalence of anti-BDV-p24 antibody was 20.73% (17/82), significantly higher than that of the controls (0%, chi(2) = 6.673, P = 0.010). The arachidonic acid level was significantly lower in the CFS group than in the controls (P > 0.05) and there were no differences in linoleic acid and ETA (both P > 0.05). The level of L-carnitine was 6.4336 +/- 3.4225, significantly lower than that of the control group (7.6666 +/- 3.5819, t = 2.025, P = 0.045) and the L-carnitine level was increased 2 weeks after supplementary treatment, together with improvement of symptoms. CONCLUSION: Most of the CFS patients are young and middle-aged. Descent of reorganization is common in these patients. Psychological abnormalities exist in most patients. Some patients are infected with BDV, some with deficiency of nutrition and/or abnormality of energy metabolism.

[Cognitive function and psychological characteristics of patients with chronic fatigue syndrome].

OBJECTIVE: To investigate the cognitive function and psychological characteristics of the patients with chronic fatigue syndrome (CFS) in China and analyze its relation with primary psychological diseases. METHODS: Ninety-one patients with CFS who visited the People's Hospital, Peking University, in Beijing from Beijing, Shanghai, Tianjin, Heilongjiang, Jilin, Hebei, Inner Mongolia, Shanxi, Shandong, Sichuan, Gansu, Fujian, and Guangdong, 42 males and 49 females, aged 37 +/- 7, 43% of which had the record of formal schooling of regular college course or over and 21 of which had the record of formal schooling of college for professional training, and 58% of which showed clear causes, diagnosed by the CDC criteria 1994, underwent case history collection, physical examination, necessary laboratory test, memory test, and SCL-90, Hamilton depression rating scale (HAMD), and Hamilton anxiety rating scale (HAMA) testing. Thirty healthy persons, 14 males and 16 females, aged 37 +/- 7, were used as controls., A table of case file was established based on the CDC criteria 1994 for each patient to record the relevant data. Independent-Samples T Test was used to compare the memory quotient, the total score and general mean score of SCL-90, the score of HAMD and HAMA. Analyzed the impairment of cognitive function and psychological characteristics of patients with CFS. RESULTS: The most common symptoms was descent of remembrance and/or attention (82/91, 90%). The memory quotient of the CFS patients was 85 +/- 14, significantly lower than that of the healthy controls (98 +/- 12, t = 4.627, P = 0.000). The total score of SCL-90 of the CFS patients was 192 +/- 47, significantly higher than that of the healthy controls (140 +/- 46, t = 5.297, P = 0.000). The symptoms with a factor score > or = 2.0 in SCL-90 included obsessive-compulsive symptoms (61/91, 67%), somatization (61/91, 67 %), depression (57/91, 63%), and anxiety (49/91, 54%). The HAMD score of the CFS patients was 9.9 +/- 6.1, significantly higher than that of the healthy controls (6.5 +/- 2.5, t = 2.948, P = 0.004). The HAMA score of the CFS patients was 9.9 +/- 7.0, significantly higher than that of the healthy controls (5.9 +/- 2.9, t = 3.015, P = 0.003). CONCLUSION: The CFS patients in China have an obvious impairment of remembrance and show different psychological abnormalities that are different from those of the patients with primary psychological diseases.

[Curative effect of gabapentin on refractory epilepsy].

OBJECTIVE: To assess the safety and efficacy of gabapentin in treatment of refractory epilepsy. METHODS: Sixty-six patients with refractory epilepsy were treated with gabapentin 200 mg/d and 72 patients with placebo, totally 138 patients in five hospitals in different cities in China. Double-blind study was performed to observe the times of seizure, and Mini-Mental State Examination (MMSE) and Activities of Daily Life (ADL) assessment were conducted every 4 weeks. RESULTS: In comparison with the control group, the seizure times at any time point in the GB group all decreased with significant differences at the 12th and 20th weeks. The significant efficacy rates, with the seizure times decreasing by more than 75%, in the gabapentin group were higher than those in the control group, with significant differences in the 4, 8, 16, and 20th weeks. Both the MMSE scores of the 2 groups were raised with a significant difference between the 2 groups at the 16 weeks. There was no significant difference in ADL between these 2 groups. No serious side effect was found in these 2 groups. CONCLUSION: Gabapentin at a dosage of 1200 mg/d is safe and effective in treatment of epilepsy.

[Construction and expression of the fusion protein DT389-hIL-13 and its cytotoxicity to glioma cell lines].

OBJECTIVE: To construct a fusion protein toxin DT389-hIL-13 which comprises the N-terminal 389 amino acids of diphtheria toxin (DT389) and human interlukin 13 (hIL-13), and to explore its cytotoxicity on U251 glioma cells. METHODS: The cDNA of hIL-13 gene was amplified by PCR and linked with the 3'-terminus of the gene encoding the N-terminal 389 amino acids, which correspond to the enzyme domain and transmembrane domain of diphtheria toxin. The tandem constructed gene was then inserted into an E. coli expression vector pET30a. The resulted expression vector was transformed into E. coli BL21 and induced by IPTG. The expressed protein was analyzed by SDS-PAGE and Western blot analysis. U(251) glioma cells were cultured DT389-hIl-13 was added into the culture. The cytotoxicity was determined using colorimetric MTS proliferation assay. RESULTS: The expression plasmid pET30a/DT389-hIL13 was constructed with correct sequence. The recombinant protein was successfully expressed in E. coli in manner of inclusion body and with a relative molecular weight of about 55 000, which reacted well with both anti-diphtheria toxin and anti-hIL-13 polyclonal antibody in Western blot assay. The purified recombinant chimeric toxin was found to effectively inhibit the prolifieration of glioblastoma multiforme cells bearing high affinity hIL-13 receptors, and resulted in dose-dependent relationship with 50% inhibition concentration (IC(50)) of 5 x 10(-)11mol/L. CONCLUSION: Prokaryotic expression system can be recruited to produce recombinant chimeric toxin DT389-hIL-13. The results may lay a foundation for preparing specific the agent targets for tumors overexpressing IL-13 receptor.