Detection and In-house Quality Control Assessment for Cyclosporin A and Tacrolimus by High Performance Liquid Chromatography-tandem Mass Spectormetry Method / 医药导报

Objective To establish a quality control method for monitoring the blood concentrations of cyclosporin A and tacrolimus by HPLC-MS/MS,and to evaluate the quality control samples using the Westgard multi-rule theory.Methods HPLC-MS/MS was used to determine the concentration of cyclosporin A and tacrolimus in human whole blood.The quality control samples of low,medium and high concentration levels in the therapeutic drug monitoring process were statistically analyzed,Levery-Jennings and Z-score quality control charts were drawn,and the Westgard multi-rule theory was applied for in-house quality control evaluation.Results The established method was fully validated with linear ranges of 10.40-1 040.00 ng·mL-1 and 0.50-49.50 ng·mL-1,the quantification limits were 10.40 and 0.50 ng·mL-1,respectively.The extraction recoveries were 108.61％-113.24％and 101.99％-109.37％,respectively.The matrix factors normalized by internal standard were 106.68％-111.27％and 95.70％-97.81％for cyclosporin A and tacrolimus,respectively.The intra-day and inter-day accuracy and precision were less than 15.0％.Other parameters were also validated and met the acceptance criteria.Levery-Jennings and Z-score quality control charts showed that there were 4 warnings(violation of the 12s rule)in the results of the 26 groups of quality control samples in the third quarter of 2022,and no phenomenon was observed to be out of control.Conclusion The established in-house quality control system for therapeutic drug monitoring of cyclosporin A and tacrolimus can effectively ensure the accuracy of blood drug concentration detection.

Prognostic model on pregnancy outcomes for women with recurrent spontaneous abortions treated with cyclosporin A: A single-institution experience

Abstract Background: This study aimed to identify prognostic factors for pregnancy outcomes and construct a prognostic model for pregnancy outcomes in women with Recurrent Spontaneous Abortions (RSA) treated with cyclosporin A. Methods: A total of 154 RSA patients treated with cyclosporin A between October 2016 and October 2018 were retrospectively recruited. Multivariate logistic regression was applied to identify the prognostic factors for pregnancy success in RSA women treated with cyclosporin A. The Receiver Operating Characteristic (ROC) curve was applied to construct prognostic value, and the prognostic performance was assessed using area under the ROC. Results: After adjusting potential confounding factors, the authors noted increased age (OR = 0.771; 95 % CI 0.693‒0.858; p < 0.001) and positive antinuclear antibodies (OR = 0.204; 95 % CI 0.079‒0.526; p = 0.001) were associated with a reduced incidence of pregnancy success, while positive anti-β2 glycoprotein-I-antibody (OR = 21.941; 95 % CI 1.176‒409.281; p = 0.039) was associated with an increased incidence of pregnancy success after treated with cyclosporin A. The AUC of combining these variables for predicting pregnancy failure was 0.809 (95 % CI 0.735‒0.880). Conclusion: This study systematically identified the prognostic factors for pregnancy success in women treated with cyclosporin A, and the constructed prognostic model based on these factors with relatively higher prognostic value. Further large-scale prospective studies should be performed to validate the prognostic value of the constructed model.

Mechanistic study of Cyclosporin A in alleviating cardiac injury in model mice with acute pancreatitis / 中国免疫学杂志

Objective:To observe whether Cyclosporin A(CSA)treatment may protect acute pancreatitis(AP)mice and related cardiac injury by consequent attenuation of systemic inflammation via regulating macrophage polarization.Methods:RAW264.7 cells were used for in vitro experiments,stimulated with 0,5,10 and 20 nmol/L CSA for 24 hours,and M2 markers were detected by flow cytometry.C57BL/6J mice were used for experiments in vivo.The mice were randomly divided into 3 groups(n=15):control group,AP model group(intraperitoneal injection of L-arginine)and AP+CSA(20 mg/kg)group,CSA was administered in a pre-treated manner.ELISA kit was used to detect the indexes of pancreatic and myocardial injury in mice;HE staining was used to detect the pathological changes of pancreas and heart tissue;TUNEL method was used to detect apoptosis in tissue sections;CETSA was used to determine the relationship between CSA and PKM;The expressions of PKM2,HIF1α,p-STAT1 and p-STAT6 were detected by Western blot.Results:CSA increased the number of M2 macrophages and decreased the number of M1 macrophages in a dose-dependent manner in vitro.CSA pretreatment significantly improved pancreatic structure and myocardial injury in AP mice,decreased pancreatic histopathological score and serum amylase,lipase,TNF-α,CK-MB,LDH and cTnT levels.CSA pretreatment significantly reduced the number of TUNEL positive cells in myocardium of AP mice.Flow cytometry analysis showed that CSA pretreatment significantly inhibited the proportion of CD11c+F4/80+ and promoted the ratio of CD206+F4/80+ in AP induced myocardial macrophages.CETSA analysis showed that PKM2 was the target of CSA.Conclusion:CSA can significantly improve the severity of L-arginine-induced cardiac damage in AP model mice,and its mechanism of action is related to increasing the number of M2 macro-phages and inhibiting the production of proinflammatory cytokines.

Comparison of the inhibitory effect of topical cyclosporine A 0.1% and topical anti-VEGF application in an experimental model of corneal neovascularization / Comparação do efeito inibitório da ciclosporina A 0,1% tópica e aplicação de anti-VEGF tópica em um modelo experimental de neovascularização da córnea

ABSTRACT Purpose: The aim of this study was to compare the effects of topical cyclosporine 0.1% and bevacizumab on experimentally induced corneal neovascularization in a rat model. Methods: A total of 30 adult Sprague-Dawley rats were used in this experimental study. The central cornea of the rats was cauterized chemically. The rats were randomly enrolled into three groups as follows: Group 1 received bevacizumab 1%, Group 2 received cyclosporine 0.1%, and Group 3 received isotonic saline twice a day for 28 days. Slit-lamp examination of all rats was performed at the 3rd and 28th day. The rats were then sacrificed, and the corneas were excised. The number of blood vessels, state of inflammation, and collagen formation were evaluated histopathologically in the corneal sections. Results: Corneal opacity and edema grades were significantly lower in Group 2 than in Group 3 (p=0.04 and 0.00, respectively). In the histopathological examination, Group 2 demonstrated significantly lesser number of blood vessels than Group 3 (p=0.001). Regarding collagen formation, Group 2 exhibited more regular collagen formation than Groups 1 and 3 (p=0.03). Inflammation grades were significantly lower in Groups 1 and 2 than in Group 3 (p=0.014 and 0.001, respectively). Conclusion: Topical bevacizumab is effective in inhibiting newly formed corneal neovascularization. The topical cyclosporine 0.1% treatment appears to be more effective than the topical bevacizumab treatment.

RESUMO Objetivo: Comparar os efeitos da ciclosporina tópica 0,1% e do bevacizumabe na neovascularização da córnea produzida experimentalmente em um modelo com ratos. Métodos: Trinta ratos Sprague-Dawley adultos foram usados neste estudo experimental. A córnea central dos ratos foi cauterizada quimicamente. Os ratos foram distribuídos aleatoriamente em três grupos. O grupo 1 recebeu bevacizumabe a 1%, o grupo 2 recebeu ciclosporina tópica a 0,1% e o grupo 3 recebeu solução salina isotônica duas vezes ao dia durante 28 dias. O exame de lâmpada de fenda de todos os ratos foi realizado no terceiro e no vigésimo oitavo dias. Os ratos foram então sacrificados e as córneas excisadas. Nos cortes da córnea, o número de vasos sanguíneos, o estado de inflamação e a formação de colágeno foram avaliados em uma análise anatomopatológica. Resultados: No Grupo 2, os graus de opacidade e de edema da córnea foram significativamente menores que no Grupo 3 (p=0,04 e 0,00, respectivamente). No exame histopatológico, o Grupo 2 apresentou um número significativamente menor de vasos sanguíneos do que o Grupo 3 (p=0,001). Em relação à avaliação da formação de colágeno, esta mostrou-se mais regular no Grupo 2 que no Grupo 1 e no Grupo 3 (p=0,03). Os graus de inflamação foram significativamente menores no Grupo 1 e no Grupo 2 em comparação com o Grupo 3 (p=0,014 e 0,001, respectivamente). Conclusão: O bevacizumabe tópico é eficaz na inibição da neovascularização da córnea recém-formada. O tratamento tópico com ciclosporina a 0,1% parece ser mais eficaz em comparação ao tratamento tópico com bevacizumabe.

Research progress of CYPA in malignant tumors / 国际生物医学工程杂志

Cyclosporin A (CYPA) is an important member of the cyclophilin family, encoded by the peptidyl prolyl isomerase A gene, and has a variety of important biological functions, mainly involved in inflammation, immunity, and other pathophysiological processes. In addition, CYPA plays a regulatory role in tumor cell proliferation, invasion, apoptosis, metastasis, angiogenesis, and epithelial-mesenchymal transition through various molecular mechanisms, among which the specific binding of CYPA to CD147 has received wide attention. In this review, the mechanism of CYPA in various malignant tumors was mainly reviewed, and its regulatory mechanism and potential interventions in malignant tumors were analyzed, with the aim that CYPA will play an important role in the early diagnosis and precise treatment of tumors in the future.

Evaluation of the Efficacy of Cyclosporin A Combined with Recombined Human Erythropoietin in the Treatment of Patients with Chronic Aplastic Anemia / 中国医学科学院学报

Objective To compare the efficacy and safety of cyclosporin A(CsA)and CsA combined with recombined human erythropoietin(rhEPO)in the treatment of patients with chronic aplastic anemia(CAA).Methods Data of 79 patients with CAA treated at Department of Hematology,PUMC Hospital between January 2016 and June 2018 were collected for retrospective analysis.Forty-five patients were treated with CsA+rhEPO,and the other 34 patients with CsA alone.All the enrolled patients were treated for at least 1.5-2.0 years and followed for at least 1.0 year.The efficacy,side effects,long-term outcomes were compared between the two groups,and factors that may influence the efficacy were analyzed.Results The patients treated with CsA+rhEPO included 14 males and 31 females,with a median age of 43(19,73)years old.The median treatment duration of CsA and rhEPO was 26(12,38)and 4(3,6)months,respectively,and the median followed-up time was 24(12,42)months.The patients treated with CsA alone included 16 males and 18 females,with a median age of 36(16,85)years old.The median CsA treatment duration was 24(12,40)months and the median follow-up time was 25(12,40)months.There was no statistical difference in baseline characteristics between the two groups(all

Therapeutic drug monitoring in the individualized administration of cyclosporin A: Application and research progress / 中国临床药理学与治疗学

Cyclosporine A (CsA) is a kind of cellular immunosuppressant, which is widely used in organ transplantation, blood diseases and autoimmune diseases. Because of its poor oral bioavailability, individual differences and prone to adverse reactions, so clinical therapeutic drug monitoring (TDM) and individual administration of CsA can ensure its safety and effectiveness. However, the treatment window of CsA is narrow, and its blood concentration is affected by age, sex, diet, drug factors, genetic factors and so on. Therefore, combined with the literature reports at home and abroad, this paper reviews the research on the application of TDM and individual drug administration of CsA, in order to provide more valuable reference for clinical safe and rational drug use.

<i>In vitro</i> interactions between antifungals and Cyclosporin A against Fusarium solani / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the<i> in vitro</i> interaction between antifungals and tacrolimus acting alone or in combination against Fusarium solani.<p>METHODS: According to Clinical and Laboratory Standards Institute(CLSI)M27-Ed4 and M38-A3, 22 strains of Fusarium solani were used to perform drug sensitivity tests with chessboard microdilution method by cyclosporin A combined with 4 kinds of antifungal drugs <i>in vitro</i>.<p>RESULTS: The MIC ranges of natamycin, voriconazole, amphotericin B and fluconazole against 22 strains of Fusarium solani were 2-8, 1-8, 1-8 and 8-512μg/mL respectively. When combined with tacrolimus <i>in vitro</i>, the synergistic effects of fluconazole and Amphotericin B were observed in 64% and 41% strains respectively. There were no antagonistic effects observed in all combined drug tests. With the combination, the sensitivity of Fusarium to amphotericin B was significantly increased from 4.5% to 68.2%(<i>P</i><0.001).<p>CONCLUSION: Fusarium solani is sensitive to natamycin <i>in vitro</i> and is partially sensitive to voriconazole. When combined with cyclosporine A, it can produce synergistic effects with fluconazole and amphotericin B, and significantly increase the sensitivity of Fusarium solani to amphotericin B drugs.

The protective effects of cyclosporin A on vascular permeability in sepsis rats / 药学学报

The protective effects of cyclosporin A (CsA), an inhibitor of mitochondrial permeability transition pore (MPTP), on vascular permeability in sepsis rats were investigated. Cecal ligation and puncture (CLP)-induced sepsis rats were used for in vivo studies, and the effects of CsA (1 and 5 mg·kg-1) on vascular permeability of lung, kidney, and intestine, mitochondrial respiratory control ratio, and the survival of the sepsis rats were observed. Lipopolysaccharide (LPS) was used for stimulating vascular endothelial cells (VECs) in vitro, and the effects of CsA on leakage of microvascular, immunofluorescence of zonula occludes-1 (ZO-1), and transendothelial electrical resistance (TER) were observed. All the animal welfare and experimental procedures are in accordance with the regulations of the Animal Ethics Committee of the Army Medical University. Compared with sham-operated group, the vascular permeability of lung, kidney, and intestine in sepsis rats increased significantly (P<0.05). Compared with conventional treatment group, CsA could significantly decrease the vascular permeability of lung, kidney, and intestine (P<0.05 or P<0.01), and prolong the survival period. The results of microcirculation also showed that CsA could significantly reduce the permeability of mesenteric venules in sepsis rats. At the cellular level, LPS stimulation significantly increased the permeability of vascular endothelial cells, including the decrease of transmembrane resistance and protein expression of ZO-1 (P<0.05). CsA can significantly reduce the increase of permeability of vascular endothelial cells induced by LPS stimulation (P<0.01). The function of mitochondria in the kidneys and intestines of sepsis rats was obviously impaired, and the respiratory control ratio of mitochondria was decreased. LPS significantly increased MPTP opening of VECs, while CsA significantly inhibited MPTP opening and improved mitochondrial function. CsA may protect mitochondrial function by inhibiting the opening of MPTP and play a protective role in the vascular permeability of sepsis rats. This study will provide an insight for the treatment of sepsis vascular leakage.

Role of cyclophilin A during coronavirus replication and the antiviral activities of its inhibitors / 生物工程学报

Cyclophilin A (CypA) is a widely distributed and highly conserved protein in organisms. It has peptidyl-prolyl cis/trans isomerase activity and is a receptor for cyclosporin A (CsA). Coronaviruses are enveloped, single-stranded, positive-sense RNA viruses. Seven types of coronaviruses are currently known to infect humans, among which SARS-CoV, MERS-CoV, and SARS-CoV-2 are fatal for humans. It is well established that CypA is essential for the replication of various coronaviruses such as SARS-CoV, CoV-229E, CoV-NL63, and FCoV. Additionally, CsA and its derivatives (ALV, NIM811, etc.) have obvious inhibitory effects on a variety of coronaviruses. These results suggest that CypA is a potential antiviral target and the existing drug CsA might be used as an anti-coronavirus drug. At the end of 2019, SARS-CoV-2 raged in China, which seriously theatern human health and causes huge economic lases. In view of this, we describe the effects of CypA on the replication of coronaviruses and the antiviral activities of its inhibitors, which will provide the scientific basis and ideas for the development of antiviral drugs for SARS-CoV-2.

Effect of cyclosporine A combined with glucocorticoid in the treatment of corneal ulcer / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To study the effect of cyclosporine A combined with glucocorticoid in the treatment of corneal ulcer.<p>METHODS: From May 2015 to May 2018, 200 patients with corneal ulcer were selected and divided into combined group and glucocorticoid group according to different treatment methods, 100 cases each. In the combined group, cyclosporine A was given on the basis of the glucocorticoid group. C-reactive protein(CRP)and interleukin-6(IL-6)were measured to evaluate the quality of life, clinical symptoms, treatment efficiency, recurrence rate and incidence of adverse reactions.<p>RESULTS: After treatment, the levels of CRP and IL-6 in the two groups were lower than that before treatment, and the quality of life was higher than that before treatment(<i>P</i><0.05). The level of CRP and IL-6 in the combined group was lower than that in the glucocorticoid group, and the quality of life was higher than that in the glucocorticoid group(<i>P</i><0.05). The time of conjunctival hyperemia, ophthalmalgia and ulcer healing in the combined group were lower than those in the glucocorticoid group(<i>P</i><0.05). The effective rate of combined group was higher than that of glucocorticoid group, and the recurrence rate of combined group was lower than that of glucocorticoid group(<i>P</i><0.05).<p>CONCLUSION: Cyclosporine a combined with hormone is effective in the treatment of corneal ulcer, which can improve the clinical symptoms, reduce the inflammation, improve the quality of life and reduce the recurrence.

Agrandamiento gingival generalizado en un paciente con trasplante renal / Generalized gingival enlargement in a patient with renal transplant

Introducción: El agrandamiento gingival es el aumento exagerado y desfigurante del volumen de la encía. Su aparición se asocia a fármacos, entre los que se encuentran los inmunosupresores y los bloqueadores de los canales de calcio como la ciclosporina A y amlodipino. Objetivo: Describir un caso clínico de agrandamiento gingival asociado a ciclosporina A y amlodipino, con periodontitis crónica subyacente, su tratamiento y prevención de recidiva. Presentación del caso: Paciente masculino, de 50 años de edad, antecedentes de hipertensión arterial, asma bronquial y hepatitis C, además de presentar insuficiencia renal crónica para la cual se le realizó un trasplante renal. Recibe tratamiento con ciclosporina A y amlodipino. Al examen clínico se observaron aumento de volumen generalizado en la encía, que cubría completamente la corona de los dientes, bolsas periodontales de 5 a 8 mm, sangramiento gingival y movilidad dentaria. Principales comentarios: El proceso diagnóstico permitió comprobar que además del agrandamiento gingival generalizado existía una periodontitis crónica generalizada. Conclusiones: La ingestión de un inmunosupresor como la ciclosporina A con el uso de un bloqueador de los canales de calcio, el amlodipino, y la influencia de factores de connotación local, parecen ser los responsables de la aparición combinada del agrandamiento gingival generalizado y la periodontitis crónica concomitante. La fase higiénica contribuyó considerablemente a mejorar el estado periodontal, cuya solución definitiva se alcanzó con la cirugía periodontal convencional. Se corrobora la importancia del examen periodontal en pacientes candidatos a trasplantes de órganos(AU)

Introduction: Gingival enlargement is an exaggerated and disfiguring increase in gum volume, associating its appearance with drugs like immunosuppressants and calcium channel's blockers such as cyclosporine A and Amlodipine. Objective: To describe a clinical case of gingival enlargement associated to cyclosporine A and amlodipine, presenting chronic underlying periodontitis, its treatment and prevention in case of recurrence. Case Presentation: Male patient, 50 years old with a history of arterial hypertension, bronchial asthma and hepatitis C, and presenting chronic renal failure leading renal transplant. The patient was treated with cyclosporine A and amlodipine. In the clinical examination was observed an increased volume in the gum, which completely covered the crown of the teeth, also periodontal bags of 5 to 8 mm, gingival bleeding and dental mobility. Main Comments: The diagnostic process allowed to verify that in addition to the generalized gingival enlargement there was a generalized chronic periodontitis. Conclusions: The ingestion of an immunosuppressant such as Cyclosporin A with the use of a calcium channel's blocker, amlodipine, and the influence of local connotation factors seem to be responsible for the combined appearance of generalized gingival enlargement and concomitant chronic periodontitis. The hygienic phase contributed considerably to improve the periodontal state, whose definitive solution was achieved with conventional periodontal surgery. The importance of periodontal examination in patients who are candidates for organ transplants is corroborated(AU)

Assessment of The Potential Role of Parsley (Petroselinum Crispum) Leaves Extract in Ameliorating Cyclosporin A- Induced Nephrotoxicity in Rats

In this study, a potentiometric titration method by Calvin-Bjerrum and Irwing-Rosotti was used to investigate binarycomplexes of ibandronate sodium, a nitrogen-containing bisphosphonate, with Ca(II), Mg(II) and Sr(II). Dissociationconstants (pKa) of ibandronate sodium were measured and the stability constants of the complexes formed in aqueoussolutions at 22 oC (I = 0.11 M NaClO4) were determined. The stoichiometry of ibandronate sodium/metal complexes wasfound as 1/1 for each metal ion.

Simvasatin plus cyclosporin A inhibits obliterative bronchiolitis in a murine heterotopic tracheal transplant model / 中华器官移植杂志

Objective@#To explore the effect of simvastatin combined with cyclosporin A treatment on the development of obliterative bronchiolitis in a murine heterotopic tracheal transplantation model.@*Methods@#Murine tracheals were heterotopically transplanted from BALB/c donors into C57BL/6 recipients. Transplanted animals received either control chow, chow containing simvastatin, chow containing cyclosporine A, or chow containing simvastatin and cyclosporine A. beginning immediately after transplantation. Epithelial loss and luminal obstruction were analyzed by morphometry. Immunohistochemistry assay was used for quantifying inflammatory cell infiltration and expression of chemokine in tracheal allografts. collagen deposition was studied by picro sirius red staining.Group t test was used to calculate the difference between groups.@*Results@#simvastatin combined with cyclosporin A treatment reduced chemokine(MCP-1, RANTES)release, inhibited CD4+ and CD8+ T cells and macrophages accumulation in tracheal allografts, resulting in limited bronchial inflammation and diminished epithelial loss. simvastatin plus cyclosporin A treatment also inhibited proliferation of myofibroblast cells, reduced MMP-2 release and decreased the amounts of type I and III collagen deposition, resulting in preserved luminal patency and inhibited development of OB compared with those of controls.@*Conclusions@#When simvastatin was used in combination with CsA, the development of OB was significantly inhibited.

Performance Evaluation of the ARCHITECT i2000 for the Determination of Whole Blood Cyclosporin A and Tacrolimus

Maintaining immunosuppressant concentrations within the therapeutic range in organ recipients requires regular monitoring. The blood concentrations of immunosuppressants are routinely measured using one of several automated immunoassays, such as chemiluminescence immunoassays (CLIAs) and liquid chromatography-tandem mass spectrometry (LC-TMS). The ARCHITECT i2000 immunoassay analyzer (Abbott Diagnostics, USA) was developed as an automated CLIA analyzer for the measurement of cyclosporin A and tacrolimus in whole blood. Here, the precision and linearity of the ARCHITECT i2000 analyzer for the detection of cyclosporin A and tacrolimus in whole blood were evaluated according to Clinical and Laboratory Standards Institute guidelines and were compared with those of an LC-TMS detection method. The total coefficient of variation for the two drugs was less than 10%, and they showed linearity values of 0.97 or more, which was within the manufacturer's range. The measurements of both immunosuppressants by the ARCHITECT i2000 were closely correlated with measurements determined by LC-TMS. However, most measurements were lower with LC-TMS than with the ARCHITECT i2000. Measurement of cyclosporin A and tacrolimus in whole blood using the ARCHITECT i2000 showed very satisfactory performance in terms of precision and linearity as well as good correlation with the comparative method.

Identification of the related substances of cyclosporin A by LC-MS techniques / 中国药科大学学报

@#To identify the related substances of cyclosporin A by LC-MS techniques, the separation of cyclosporin A and its related substances was carried out on a Hypersil BDS C18(100 mm×4. 6 mm, 2. 4 μm)column with isocratic elution by a mixture of acetonitrile-water-MTBE and formic acid(430 ∶520 ∶50 ∶1)as the mobile phase. Cyclosporin A and its 29 related substances(9 process related and 20 degradants)were well separated under the established conditions. Among them, 13 were listed in EP and the rest 16 were unknown products having not been reported before. Electrospray positive ionization high resolution TOF/MS was used for the determination of the accurate mass and elemental composition of parent ions of all the components, and triple quadrupoles tandem mass was employed for the product mass spectra determination. Thence, the structures of all the 29 detected substances were successfully characterized through spectra elucidation and the fragmentation pathways analysis. The established LC-MS method was successfully employed for the separation and identification of the related substances of cyclosporin A and it is useful for its fermentation processes and quality control.

Molecular Pathway of Psoralidin-Induced Apoptosis in HepG2 Cell Line / 中国结合医学杂志

OBJECTIVE@#To test the role of psoralidin in human liver cancer HepG2 cells in vitro.@*METHODS@#Cell viability was assessed by methylthiazolyldiphenyl-tetrazolum bromide assay and apoptotic cells were labeled by annexin V then sorted by flow cytometry. Protein expressions of caspase-3, caspase-8, caspase-9, Bax, Bid, Bcl-2, Bcl-xL and p53 were examined by western blot while activity of caspase-3, -8 and -9 were also determined.@*RESULTS@#Psoralidin reduces cell viability greatly in a time dependent manner (64%, 40%, 21%, 12% at 2, 6, 24 and 48 h treatment with 64 μmol/L psoralidin respectively) and up-regulates activities of caspase-3, -8 and -9 in a concentration dependent manner (between 4 to 64 μmol/L). Psoralidin also increases the expression of pro-apoptosis genes Bax, Bid and p53 while decreases the expression of pro-survival genes Bcl-2 and Bcl-xL, both in a concentration dependent manner between 4 and 64 μmol/L (P<0.05 at 16 and 64 μmol/L). Caspase-3 inhibitor (Ac-DEVD-CHO at concentrations between 10 to 20 μmol/L), p53 inhibitor (pifithrin-α at 5 μmol/L) and cyclosporin A can attenuate the apoptotic effect of psoralidin.@*CONCLUSION@#The cytotoxic role of psoralidin might work through both intrinsic and extrinsic apoptotic pathway.

Simvasatin plus cyclosporin A inhibits obliterative bronchiolitis in a murine heterotopic tracheal transplant model / 中华器官移植杂志

Objective To explore the effect of simvastatin combined with cyclosporin A treatment on the development of obliterative bronchiolitis in a murine heterotopic tracheal transplantation model .Methods Murine tracheals were heterotopically transplanted from BALB/c donors into C57BL/6 recipients .Transplanted animals received either control chow ,chow containing simvastatin ,chow containing cyclosporine A ,or chow containing simvastatin and cyclosporine A . beginning immediately after transplantation .Epithelial loss and luminal obstruction were analyzed by morphometry .Immunohistochemistry assay was used for quantifying inflammatory cell infiltration and expression of chemokine in tracheal allografts .collagen deposition was studied by picro sirius red staining .Group t test was used to calculate the difference between groups .Results simvastatin combined with cyclosporin A treatment reduced chemokine (MCP-1 , RANTES ) release , inhibited CD4+ and CD8+ T cells and macrophages accumulation in tracheal allografts ,resulting in limited bronchial inflammation and diminished epithelial loss .simvastatin plus cyclosporin A treatment also inhibited proliferation of myofibroblast cells ,reduced M M P-2 release and decreased the amounts of type I and III collagen deposition ,resulting in preserved luminal patency and inhibited development of OB compared with those of controls .Conclusions When simvastatin was used in combination with CsA ,the development of OB was significantly inhibited .

The protective effect of Xuebijing on mouse model of immunosuppressed sepsis / 天津医药

@#Objective To observe the effect of Xuebijing on immunosuppressed sepsis of model mice. Methods The 152 mice were randomly divided into control group (Control), immunosuppression group (IM), immunosuppressed sepsis model group (ISM) and Xuebijing treatment group (XT). There were thirty – eight mice for each group. For group IM, cyclosporine A was injected intraperitoneally along the median line of the lower abdomen for immunosuppression, 25 mg/kg， 1 time every other day for a total of 3 times. For group ISM, after immunosuppression, 300 μL escherichia coli 44102 with a concentration of 1×109 CFU/mL was injected intraperitoneally along the midline of the lower abdomen. The 30 minutes after the establishment of the immunosuppressive sepsis model, Xuebijing 4 mL/kg was intraperitoneally injected along the midline of the lower abdomen in the ISM group. After 30 minutes, the same dose of Xuebijing injection was repeated once. Control group was injected intraperitoneally with the same amount of normal saline. (1) After 8 h, 4 mice in each group were taken for blood bacterial culture. (2) After 12 h, 10 mice in each group were taken for detecting CD3+CD4+ and CD3+CD8+ in peripheral blood using flow cytometry. (3) After 12 h, 10 mice in each group were taken for detecting blood levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) by enzyme-linked immunosorbent assay (ELISA). (4) After 12 h, 4 mice in each group were taken for detecting high-mobility group protein (HMGB1) by Western blot assay. (5) After 12 h, 10 mice in each group were taken for detecting alanine aminotransferase (ALT), aspartate aminotransferase (AST), urea nitrogen (BUN) and serum creatinine (CR) levels by automatic biochemical analyzer. Results Compared with the ISM group, the ratio of CD3+CD4+/CD3+CD8+ was significantly increased, the number of blood bacteria culture decreased obviously in the XT group. Liver and kidney function indicators ALT, AST, CR, BUN and inflammatory factor indicators TNF-α, IL-6 and HMGB1 were also significantly decreased in the XT group. Conclusion Xuebijing can obviously modulate the immunosuppression, against bacteria, inhibit the inflammatory reaction, and protect the vital organs.

Pharmaceutical Care of Hypertension Induced by the Combination of Cyclosporine A and Voriconazole in A Child with Aplastic Anemia / 医药导报

Objective To investigate the pharmacy practice of clinical pharmacists' attendance in the treatment of adverse reactions caused by drug interactions. Methods Analysis was conducted on the causes and treatment of hypertension in one pedinatric patient with aplastic anemia induced by combined use of cyclosporine and voriconazole. Results Cyclosporine A could lead to prolonged hypertension.Cyclosporine A dosage should be adjusted;antihypertensive drugs should be used in necessity.When choosing antihypertensive drugs,drug interaction with cyclosporine A should be considered. Conclusion Clinical pharmacists could manage adverse drug reactions by analyzing drug interactions,and offer suggestions for ADR patients.