AIM　Diclofenac potassium sustained-release tablets were developed as a gel matrix for using twice daily and evaluated in vitro release characteristics. METHODS　The formulations were screened according to the designed in vitro release rate by the use of HPMC as the gel matrix and the hydrophobic retarding agent to modify the release. RESULTS　The release was properly characterized by the diffusion mechanism, influenced by pH of the media, slightly affected by the basket rotating speed, without the influences from the pressure exerted during the tabletting procedure. CONCLUSION　The selected formulation of diclofenac potassium sustained-release tablets could ensure the desired in vitro release rate. In addition, it proved a good manufacturing reproducibility.

Audiometry, Speech ; Humans ; Language

Objective Liver metastasis model of human primary gastric lymphoma in nude mice was reproduced for an experiment to evaluate the inhibitory effect of cytokine-induced killer (CIK) cells on the growth and liver metastasis of primary gastric lymphoma. Methods Surgical orthotopic implantation of a histologically intact liver metastasis fragment derived from a surgical specimen of a patient with metastatic gastric lymphoma was initally implanted, in order to reprodueing a liver metastasis model of human primary gastric lymphoma. Peripheral blood mononuclear cells (PBMC), isolated by blood cell separator from healthy donors and patients with primary gastric lymphoma, were incubated in vitro. rhIFN-?, rhIL-2 and anti-CD3 McAb were added to PBMC in order to prepare CIK cells as well as lymphokine-activated killer (LAK) cells. CIK and LAK cells from different donors were used in treating gastric malignant lymphoma, so as to investigate the inhibitory effect of 2 kinds of effector cells on the growth of the tumor and liver metastasis. Results The liver metastasis model of human primary gastric malignant lymphoma in nude mice was successfully reproduced. After administration of different agents continuously for 20 days (0.3ml/d), the inhibitory rates of the following 4 groups, healthy donors LAK group (2?1010/ml), healthy donors CIK group (2?1010/ml), patients CIK group (1?1010/ml) and patient CIK group (2?1010/ml), were 39.28%, 53.57%, 40.38% and 56.42%, respectively. The liver metastasis rates in control group, healthy donors LAK group, healthy donors CIK group, patients CIK group (1?1010/ml) and patient CIK group (2?1010/ml), were 100.0%, 62.5%, 50.0%, 62.5% and 37.5%, respectively. Tumor weights of all treatment groups were lighter than that of saline group (P

Objective To investigate the effect of matrix metalloproteinase(MMP-2) in the attack of exprimental autoimmune myositis(EAM) rats, and the effects of methylprednisolone on expressions of gene mRNA and protein of MMP-2. Methods The expression levels of MMP-2 in the peripheral blood, spleen lymphocytes and muscles were detected by using RT-PCR and immunohistochemistry techniques and was compared among EAM group,EAM with methylprednisolone treatment group (EAMM) and control group. Results ⑴ The degree of episode in EAMM group was lower than that of EAM group , and the infiltration of the inflammatory cells and the necrosis of the muscles were also mild in EAMM group. ⑵There was significant upregulation of the expression of the MMP-2 gene mRNA in the lymphocytes of peripheral bood and spleen of EAM rats as compared with that of the control,Elevation of the expression of MMP-2 protein in rats muscles tissues of EAMM group was observed obviously. However, the expression of the mRNA and the protein of MMP-2 was suppressed significantly in EAM group compared with control group. Conclusions The upregulation of the expression of MMP-2 may be correlated with the onset of EAM. Methylprednisolone may relieve the degree of the pathology of EAM by the downregulation of MMP-2 .

Objective　To obtain mammalian cell expression vector of human CD154 gene. Methods　A 820 bp cDNA fragment was amplified by RT-PCR method from total RNA of human peripheral blood mononuclear cell（PBMC） activated with 10 ng／mL PMA and 1 μg／mL PHA for 8 hours. The fragment was cloned into pcDNA3.1（＋） plasmids.The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes BamH Ⅰ and EcoR Ⅰ and sequenced by Sangers-dideory-mediated chain termination. Results　This cDNA fragment included 820 bp entire coding region and a part of the 3 non-coding region. The recombinant mammalian cell expression vector of pcDNA3.1（＋）／hCD154 was constructed， the sequence of the insert was identical to the published sequence encoding human CD154 antigen. Conclusion　The recombinant mammalian cell expression vector of pcDNA3.1（＋）／hCD154 was successfully constructed.

AIM:To express recombinant hCD154-GST fusion protein, to prepare anti-hCD154 monoclonal antibody, and to investigate the effect of anti-hCD154 monoclonal antibody on graft rejection. METHODS AND RESULTS: Total RNA was prepared from human peripheral blood mononuclear cell (PBMC) activated with 10ng/mL PMA and 1 ?g/mL PHA for 8h, the total RNA was reversetranscribed to cDNA. The entire coding region and a part of the 3'non-coding regions were amplified by PCR using a pair of primers designed and synthesized according to the sequence of human CD154 gene from gene bank. The amplified product, a 820bp DNA fragment was cloned into pGEX-4T-1 plasmid expressing glutathione S-transferase(GST). The cloned insert was identified by double digestion of the cloned pGEX-4T-1 plasmid with retriction enzymes BamHⅠand EcoRⅠ.The fusion protein expression plasmid of PGEX-4T-1/hCD154 was constructed, then transformed to E coli BL21. The human CD154-GST fusion protein expression was induced by IPTG in BL21. The expression of recombinant 26kD GST and 55kD human CD154-GST fusion protein were confirmed by SDS-PAGE. CONCLUSION: We have express the recombinant human CD154-GST fusion protein. The expressed hCD154-GST fusion protein will be used to prepare anti-hCD154 monoclonal antibody, to investigate the role of anti-CD154 monoclonal antibody on graft rejection.

AIM:To obtain the light chain region(VL) and heavy chain region(VH) genes from the hybridoma cell line and analyse their sequence for construction of the engineering antibody against hCD154. METHODS: In this research ,total RNA was extracted from the hybridoma cell line 7E8, which secretes McAb against hCD154, and subjected to reverse transcription. The VL gene and VH gene were amplified by PCR, cloned into puc18 vector and sequenced by Sanger's dideoxymediated chain-termination method. RESULTS: The VL cDNA of 7E8 McAb consists of 341 bp encoding 113 amino acid residues. Compared with mouse Ig database, the VL region is in accord with the characterization of DNA sequence present in the mouse Ig Vk region , it belong to mouse V?2 light chain. The VH cDNA of 7E8 McAb consists of 354 bp encoding 118 amino acid residues. Compared with mouse Ig database, the VH region is in accord with the characterization of DNA sequence present in the mouse Ig VH region. CONCLUSION: The DNA squence analysis showed that the cloned genes code the light and heavy chain variable region of mouse respectively.

Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; complications ; Male ; Mucocutaneous Lymph Node Syndrome ; complications

Objective To study the clinical features of the variants of benign childhood epilepsy with central temporal spikes (BECT).Methods The clinical data of 12 hospitalized pediatric patients with BECT from Jan 2007 to Jan 2014 were retrospectively reviewed. Results There were 7 boys and 5 girls in 12 patients. The age of onset was from 3 to 9 years old. Two cases were dizygotic twins. The atypical symptoms included atypical absence of 10 cases, negative myoclonic seizure of 8 cases, speech expression disorders and oral-pharynx apraxia of 4 cases. The electroencephalography (EEG) of all 12 patients showed abundance of spike and waves (SW) in rolandic areas during wake-up and sleep. The SW index was 50%-85% during slow sleep in all patients.Conclusions The variants of BECT are often associated with EEG deterioration. Understanding the clinical featuress and EEG characteristics can help the diagnosis of BECT variants.

Objective The study was conducted to investigate the acute phase of convulsion related problem on the clinical manifestations,imaging and electroencephalograph (EEG) examination of purulent meningitis.Methods Cluster sampling method was employed to select children in our hospital,a total of 301 cases with purulent meningitis was analyzed retrospectively.Among them,62 cases had convulsion.The incidence of convulsion in the acute phase of the purulent meningitis,risk factors,and prognosis were analyzed.Results The convulsion incidence rate of acute purulent meningitis was 20.60％.The partial seizure was eight cases (12.90％).The secondarily generalized seizure following partial seizure was 15 cases (24.19％).The generalized seizure was 32 cases (51.61％).The convulsive status was 7 cases (11.29％).The EEG abnormality was significantly different between the convulsion group and the no convulsion group (P ＜ 0.05).The incidence of brain organic damage was significantly different between two groups (P ＜0.05).The multivariate unconditional logistic regression analysis showed,cause of disease,first symptom,disturbance of consciousness,obvious signs,and cerebrospinal fluid culture with convulsion were the relevant factors (P ＜ 0.01).Conclusions The most common seizure of purulent meningitis was the generalized seizure.Brain organic damage easily resulted in convulsion of purulent meningitis.The days of hospitalization,cause of disease,first symptom,disturbance of consciousness,obvious signs,and cerebrospinal fluid culture with convulsion were the positively relevant factors.Those positively relevant factors in combination of the clinical manifestations,imaging,and EEG examination in children would play an important role in diagnosis,treatment,and prognosis evaluation of convulsion derived from purulent meningitis.Moreover,convulsion affects the disease recovery in children with purulent meningitis.