Safety of Tacrolimus in Autoimmune Disease: Results From Post-marketing Surveillance in South Korea

Objective@#Tacrolimus, a macrolide immunosuppressant, is approved in Korea for the treatment of rheumatoid arthritis (RA), lupus nephritis (LN) and myasthenia gravis (MG). We report three prospective post-marketing surveillance studies of tacrolimus conducted in South Korea in these indications. @*Methods@#Studies were conducted according to South Korean Ministry of Food and Drug Safety requirements. Patients were followed up for the duration of the study (up to 4 years) or until treatment discontinuation. Occurrence and likely relationship with tacrolimus of adverse events (AEs), adverse drug reactions (ADRs; defined as AEs where causal relationship to tacrolimus could not be excluded) and serious AEs were recorded. Association of AEs with demographic and medical factors was evaluated by multivariable analysis. @*Results@#The studies included 740 (RA), 307 (LN) and 104 (MG) patients. The incidence of AEs was 12.7% in RA (64.2% of AEs potentially related to tacrolimus), 20.9% (37.8% potentially related) in LN and 29.8% (56.8% potentially related) in MG. The incidence of ADRs was 8.4%, 9.8% and 20.2%, respectively. Serious AEs were reported in 0.7%, 7.2% and 8.7%, respectively. The most common AEs were abdominal pain (RA), pharyngitis (LN) and diarrhea (MG). Unexpected AEs occurred in 3.5% of patients with RA, 2.9% in LN and 8.7% in MG; no pattern of unexpected AEs was apparent. Multivariable analysis demonstrated that patients with comorbidity had higher probability of experiencing an AE in RA and MG studies. @*Conclusion@#The incidence of AEs and the safety profile of tacrolimus in each indication was consistent with previous reports.

Safety of Tacrolimus in Autoimmune Disease: Results From Post-marketing Surveillance in South Korea

Objective@#Tacrolimus, a macrolide immunosuppressant, is approved in Korea for the treatment of rheumatoid arthritis (RA), lupus nephritis (LN) and myasthenia gravis (MG). We report three prospective post-marketing surveillance studies of tacrolimus conducted in South Korea in these indications. @*Methods@#Studies were conducted according to South Korean Ministry of Food and Drug Safety requirements. Patients were followed up for the duration of the study (up to 4 years) or until treatment discontinuation. Occurrence and likely relationship with tacrolimus of adverse events (AEs), adverse drug reactions (ADRs; defined as AEs where causal relationship to tacrolimus could not be excluded) and serious AEs were recorded. Association of AEs with demographic and medical factors was evaluated by multivariable analysis. @*Results@#The studies included 740 (RA), 307 (LN) and 104 (MG) patients. The incidence of AEs was 12.7% in RA (64.2% of AEs potentially related to tacrolimus), 20.9% (37.8% potentially related) in LN and 29.8% (56.8% potentially related) in MG. The incidence of ADRs was 8.4%, 9.8% and 20.2%, respectively. Serious AEs were reported in 0.7%, 7.2% and 8.7%, respectively. The most common AEs were abdominal pain (RA), pharyngitis (LN) and diarrhea (MG). Unexpected AEs occurred in 3.5% of patients with RA, 2.9% in LN and 8.7% in MG; no pattern of unexpected AEs was apparent. Multivariable analysis demonstrated that patients with comorbidity had higher probability of experiencing an AE in RA and MG studies. @*Conclusion@#The incidence of AEs and the safety profile of tacrolimus in each indication was consistent with previous reports.

Treatment of Retrogastric Pancreatic Pseudocysts by Laparoscopic Transgastric Cystogastrostomy / 华中科技大学学报（医学）（英德文版）

This paper discusses variations of laparoscopic transgastric cystogastrostomy in management of retrogastric pancreatic pseudocysts for 8 patients with symptom or pseudocysts (larger than 6 cm) companied with clinical manifestations.Using a Harmonic scalpel,two 3-5-cm incisions were made in the anterior and posterior gastric wall respectively.In the last step,the anterior gastrotomy was closed with an Endo-GIA stapler.All cases were successfully treated without large blood loss and without conversion to open surgery.The mean operative time was 114.29±19.24 min,blood loss was 157.14±78.70 mL,and mean hospital stay was 8.29±2.98 days,Gastric fistula occurred in one case on the postoperative day 7,and closed 1 month later.No bleeding was seen in all patients during the perioperative follow-up period.CT scans,given one month after the surgeries,displayed that the pancreatic pseudocysts disappeared or decreased in size,and ultrasounds showed no fluid or food residue in stomas at the third and fifth month following surgery.No patient experienced a recurrence during the follow-up period.Transgastric laparoscopic cystogastrostomy is a minimally invasive surgical procedure with a high rate of success and a low rate of recurrence,accompanied by rapid recovery.It is easy to master,safe to perform and may be the preferred option to treat retrogastric pancreatic pseudocysts.

Study on folk medicine used by Hakka in Meizhou / 中国中药杂志

Traditional or folk medicine, a multi-discipline subject, has gained much attention from all over the world at present. Based on the previous literatures, the overview on the recent advance of folk medicine used by Hakka was summarized. The key questions of folk medicine used by Hakka were also highlighted. It could be considered as a very important part of traditional Chinese medicines.

Determination of icaritin in rat plasma by HPLC-MS/MS / 药学学报

The paper is to report the development of a high-performance liquid chromatographic/tandem mass spectrometry (HPLC-MS/MS) method for the determination of icaritin (ICT) in rat plasma. After precipitated with acetonitrile from the plasma, ICT was isolated chromatographically on a Dikma C18 column. The mobile phase consisted of acetonitrile-water-acetic acid (72 : 28 : 1.5, v/v/v). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 387 --> m/z 313 and m/z 331 --> m/z 315 were used to quantify ICT and the internal standard, respectively. The linear calibration curve was obtained in the concentration range of 2.5-1,000 ng x mL(-1). The lower limit of quantification was 2.5 ng x mL(-1). The inter- and intra-day precision (RSD) were less than 9.63%, and the accuracy (relative error) was within +/-7.42%. The method was proved to be suitable for the pharmacokinetics of ICT, which offers advantages of high sensitivity and selectivity.

Prokaryotic expression of HLA-B * 2705 heavy chain and identification for its activity / 中国实验血液学杂志

In order to investigate the expression of heavy chain of HLA-B * 2705 in prokaryotic system and identify its activity, the extra-membrane gene fragment of HLA-B * 2705 was amplified from full-length HLA-B*2705 cDNA by PCR and cloned into pGEM-T vector. After identification by sequencing, the prokaryotic expressing vector pET32a (+)-B * 2705 was constructed. The antigenic activity of expressed protein was identified by Western blot and antibody blocking reaction. The results indicated that the fused HLA-B * 2705 protein expression with high efficiency was obtained. The expressed product was more than 50% of the total bacteria protein. The antigenic activity of expressed protein was confirmed by Western blot and antibody blocking reaction. It is concluded that HLA-B * 2705 fusion protein are obtained as basis for the further studies.

Zinc adsorption and desorption characteristics in root cell wall involving zinc hyperaccumulation in Sedum alfredii Hance / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Radiotracer techniques were employed to characterize (65)Zn adsorption and desorption in root-cell-wall of hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end of a 30 min short time radioisotope loading period, comparable amounts of (65)Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more (65)Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of (65)Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more (65)Zn accumulated in desorbed root-cell-wall of NHE. Zn(2+) binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn(2+) concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn(2+), whereas the desorption characteristics were different, and with most of (65)Zn binding on root of HE being available for loading into the xylem, as a result, more (65)Zn was translocated to the shoot.

A preliminary study on the expression and biological function of recombinant human SCF-TPO fusion protein / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the expression of recombinant human SCF-TPO fusion protein and its biological function.</p><p><b>METHODS</b>Four primers were designed according to known sequences of TPO and SCF. The functional amino acid domains of TPO and SCF were amplified by RT-PCR from fetus hepatocytes, respectively. The expression plasmid pET32a/SCF-TPO was constructed by VOE gene fusion technique and expressed in E. coli BL21(DE3) plysS as inclusion body after isopropyl-beta-D-1-thiogalactopyranoside (IPTG) induction. The fusion protein was tested by SDS-PAGE and Western blot. The biological functions of SCF-TPO fusion protein in MO7e cells was investigated by MTT method after purification with metal chelating chromatography.</p><p><b>RESULTS</b>The high expression SCF-TPO fusion protein was obtained, reaching up to 30% of the total cellular protein. Western blot verified the correct fusion expression and MTT results showed the growth promoting effect of the SCF-TPO fusion protein on MO7e cells, with a higher promoting activity at 100 ng/ml.</p><p><b>CONCLUSIONS</b>Expressed SCF-TPO fusion protein after renaturation has biological activity in promoting the proliferation of MO7e cells.</p>

Prediction of acute GVHD after umbilical cord blood transplantation by HLA three-dimensional structure matching / 中国实验血液学杂志

To explore a method of predicting acute graft versus host disease (aGVHD) after unrelated cord blood transplantation (UCBT), the HLA-A, -B, -DRB1 molecular three-dimensional structures in 25 patients with blood disorder who underwent UCBT and their donors were modeled by using molecular modeling technique. First, full amino acid sequences of each HLA antigen from HLA data banks were loaded down, and then amino acid sequence of extracellular antigen binding region was chosen. Third step, SPDBV software of SWISS-MODEL server was used to modeling the three-dimensional structures of each different allele of HLA-A, -B and -DRB1 between patients and donors and the parameter "root mean square deviation" (RMSD) was used to indicate the structure differences. Last, RMSD of each different HLA allele of each donor-patient pair were added together to get total RMSD. The 25 patients were divided into 3 groups: the first group did not develop aGVHD; the second group developed aGVHD graded I-II and the third group developed aGVHD graded III-IV. The results showed that in the 25 patients divided into three groups, 8 patients in the first group did not develop aGVHD (32%); 13 patients in the second group developed grade I-II of aGVHD (52%) and 4 patients in the third group developed aGVHD III-IV (16%). The total RMSDs of each group were 0.24 +/- 0.15, 0.25 +/- 0.14 and 0.47 +/- 0.22 respectively. The total RMSD of the third group was significantly higher than that of the other two groups. In conclusion, utilization of modeling HLA molecular three-dimension can predict the severe aGVHD after UCBT quickly, simply and accurately. It provides scientific basis in choosing a optimal cord blood donor to avoid severe aGVHD for physicians and the cord blood banks. And it is instructive too to direct the application of immunosuppressive agents after transplantation in clinic.

Norm study of occupational stress on the professionals in Chengdu and Chongqing area / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To establish the norm of occupational stress on the professionals in Chengdu and Chongqing area.</p><p><b>METHODS</b>According to the "Occupation Sorting Canon in the People's Republic of China", the professionals of the above-mentioned area were sorted out and randomly stratified. The normative data were derived from a sample of 2 064 participants. The modified Occupation Stress Inventory Revised Edition (OSI-R) was used to test the targets' occupational stress level (including stressors, strain and coping resources).</p><p><b>RESULTS</b>(1) In these professionals, the sex ratio was about 1:1, average age (36.4 +/- 9.4) years, average length of service (15.9 +/- 10.0) years, 63.07% people were college educated and over 80% married. (2) Descriptive statistics for OSI-R scale scores for the total normative sample, for gender samples and for occupational group samples were modulated. (3) The gender norm showed that there were heavier role overload and stronger occupational strains in males than those in females (P < 0.01). The score of personal resources in the males was not different from that in females (P > 0.05). (4) In terms of different occupations, the scores of occupational role and personal strain among various groups indicated significant differences (P < 0.01). The score of personal resources did not manifest difference (P > 0.05). (5) The major factors influencing OSI-R were sex, occupation and length of service.</p><p><b>CONCLUSION</b>The norm established can basically represent the occupational stress on professionals in Chengdu and Chongqing area.</p>

The scale and application of the norm of occupational stress on the professionals in Chengdu and Chongqing area / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To establish the scale of the norm of occupational stress on the professionals and put it into practice.</p><p><b>METHODS</b>T scores were linear transformations of raw scores, derived to have a mean of 50 and a standard deviation of 10. The scale standard of the norm was formulated in line with the principle of normal distribution.</p><p><b>RESULTS</b>(1) For the occupational role questionnaire (ORQ) and personal strain questionnaire (PSQ) scales, high scores suggested significant levels of occupational stress and psychological strain, respectively. T scores >/= 70 indicated a strong probability of maladaptive stress, debilitating strain, or both. T scores in 60 approximately 69 suggested mild levels of maladaptive stress and strain, and in 40 approximately 59 were within one standard deviation of the mean and should be interpreted as being within normal range. T scores < 40 indicated a relative absence of occupational stress or psychological strain. For the personal resources questionnaire (PRQ) scales, high scores indicated highly developed coping resources. T scores < 30 indicated a significant lack of coping resources. T scores in 30 approximately 39 suggested mild deficits in coping skills, and in 40 approximately 59 indicated average coping resources, where as higher scores (i.e., >/= 60) indicated increasingly strong coping resources. (2) This study provided raw score to T-score conversion tables for each OSI-R scale for the total normative sample as well as for gender, and several occupational groups, including professional engineer, professional health care, economic business, financial business, law, education and news. OSI-R profile forms for total normative samples, gender and occupation were also offered according to the conversion tables.</p><p><b>CONCLUSION</b>The norm of occupational stress can be used as screening tool, organizational/occupational assessment, guide to occupational choice and intervention measures.</p>

Cloning, expression and biological activity identification of a cDNA encoding the extracellular region of human b7-2 / 中国实验血液学杂志

As one important member of B7/CD28/CTLA-4 costimulatory signal pathway, B7-2 molecule plays a critical role in regulating T-cell response. In order to further explore its effects on regulation of T cell activation, proliferation and associated signal pathways, the cDNA encoding extracellular region of human B7-2 was amplified via PCR and subcloned into some prokaryotic expression vectors to express target protein in host strains. The expressed protein was identified with Western blot and MTT. Results showed that after screening, the expression level of the protein of interest attained the yield of over 20% total bacterial protein by using pGEX-4T-2 vector and E. coli BL21 (DE3)-CodonPlus-RIL host cells. The recombinant protein could specially react with B7-2 McAb and could stimulate T-cell proliferation combined with anti-CD3 antibody. In conclusion, the recombinant protein was bioactive, therefore the study will make it possible for the research of relationship between B7-2 structure and its function.

Selection of donor in mismatched hematopoietic stem cell transplantation by CREG, residue match and HLA three-dimensional structure / 中国实验血液学杂志

After search at hematopoietic stem cell banks for transplant donors, there may be several donors matched with given standards. To determine the most appropriate donor for a specific patient, the potential donors were analyzed and compared by three methods. The first is cross-reactive group (CREG) antigens, which defined as the public antigens that shared specific serological reaction patterns. The second is residue match theory, which concerned the three residues oriented upward toward the T-cell receptor. The third is comparing HLA three-dimensional structure models. The results of the three methods were not completely accorded in our case. However, some less matched donors could be excluded from the candidates and the range of selection was further reduced. It is concluded that combined application of three methods would contribute in selecting donor for hematopoietic stem cell transplantation in clinics.

Expression of alpha subunit for IL-6 receptor at mRNA and protein levels in human leukemic cells / 中国实验血液学杂志

To probe into the expression of alpha subunit for IL-6R at both mRNA and protein levels in human leukemic cells and to discuss its implication in targeted treatment for leukemia with recombinant IL-6-PE40 exotoxin fusion protein mediated by IL-6/IL-6R system, semi-quantitative RT-PCR, sequencing and FCM were used to analyze the gene and protein expression levels of alpha subunit for IL-6R in leukemic cells. Our results showed that not only mRNA but also protein of alpha subunit for IL-6R are highly expressed in the myelogenous leukemic cell lines, the relative expression levels of mRNA were KG-1(1.22) > (1.02) > U266(1.00) > U937(0.99) > HL-60(0.76). Among lymphoblastic leukemic cell lines, Raji expressed a certain amount of alpha subunit mRNA (0.77), whereas its alpha subunit protein was not detected. Expression of alpha subunit mRNA and protein were negative in lymphoblastic leukemic cell lines, HuT28 and CEM, and chronic myelocytic leukemic cell line K562. These results correlate with those of FCM highly. Noteworthily, normal human peripheral blood mononuclear cells expressed hardly protein of IL-6R alpha subunit. So this study provides sufficient experimental evidence that the targeted treatment by recombinant IL-6-PE40 can specifically kill leukemic cells highly expressing IL-6R without toxicity to normal hematopoietic cells.

Advances in research on cytokine mimetic peptides/non-peptides stimulating hematopoiesis / 中国实验血液学杂志

Cytokines such as erythropoietin (EPO) and thrombopoitein (TPO) and so on, which stimulate hematopoiesis, can regulate self-renewal, proliferation, differentiation, maturation and programmed cell death of hematopoietic cells through specifically binding to surface receptors. Recently random phage display peptide libraries and other screening methods have been used to isolate mimetic including small peptides and non-peptides molecules, which can mimic the same effects as cytokines, such as EPO and TPO, and demonstrate the similar potency and activity as EPO and TPO in a panel of in vitro biological assays and in animal experiments. These approaches are critical to further research of interactive mechanisms between cytokine and receptor, receptor activation and rational design of other desired cytokine mimetic. This review concisely introduced recent advances in research on mimetic of EPO, TPO and other cytokines and future directions.

The Prediction of the Molecular Biology Characteristics of Recombinant Fusion Exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40 / 中国实验血液学杂志

In order to confirm the reasonability of designed recombinant exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40, their molecular biology characteristics, such as flexibility, antigenicity, hydrophilicity, epitope and secondary structure, were predicted by using a computer software GOLDKEY. It had been found that the recombinant fusion exotoxin had kept the epitope characterstics of B7-1, B7-2 and PE40, and had not got new epitope, and the antigenicity in flexible linker was extxemely low. The linker inserted in the recombinant fusion exotoxin had low epitope, low antigenicity and high flexibility. Compared to B7-1, B7-2 and PE40, there are several amino acid residues changes in B7-1-Linker-PE40 and B7-2-Linker-PE40, respectively, which might have some effect on secondary structure of the recombinant fusion exotoxins. Western blot analysis revealed that both B7-1-Linker-PE40 and B7-2-Linker-PE40 could bind specifically with antibodies against B7-1, B7-2 and PE40, respectively. The result of Western blot was consistant with the computer prediction that the recombinant proteins retain the antigenicity and spacial structure of B7 and PE40. It is suggested that both fusion proteins designed and constructed were resonable and computer analysis would be helpful for us to study the biological activity of the recombinant fusion exotoxin B7-1-Linker-PE40 and B7-2-Linker-PE40 and construct other recombinant proteins further.

A New Criterion for Donor and Recipient Selection in Hematopoietic Stem Cell Transplantation - the Matching of Three-Dimensional Structure of HLA Molecular Modeling / 中国实验血液学杂志

The purpose of the research is to provide a new standard for matching of HLA three-dimensional structure, and summarize the major permissible mismatch and immunogenic mismatch antigens. The molecular modeling method was used to create HLA molecular structures by Swiss Model Server, and the comparison of the differences among the alleles was done by SPDV software with the function of iterative magic fit. The results were recorded by relative mean square deviation (RMSD, nm). The differences among alleles were scattered below 0.06 nm for HLA-A and -B molecules, and below 0.03 nm for HLA-DRB1 molecules. On the basis of the statistical analysis, when RMSD is greater than 0.04 nm for -A and -B molecules and 0.02 nm for -DRB1 molecules, the difference is meaningful and can be related with graft versus host disease. When RMSD is lower than 0.02 nm for -A and -B molecules and 0.01 nm for -DRB1 molecules, the difference is decided unmeaningful. From the data, the permissible mismatch and immunogenic mismatch alleles within HLA-A, HLA-B and HLA-DRB1 molecules were summarized. Three-dimensional structure matching is a new area in the transplantation field, much research should be done in the future.