Objective:To study the characteristics of the adverse reactions caused by tacrolimus and analyze the risk factors. Methods:Totally 280 cases of renal transplant recipients received standard treatment options were selected from 1997 to 2013 in Wuhan general hospital of Guangzhou military command. The tacrolimus-related adverse reactions and their risk factors were analyzed statistically. Results:Totally 39. 65% of the patients had the tacrolimus-related adverse reactions. The main adverse reactions were hematotoxicity,diabetes mellitus and hyperlipidemia. The univariate analysis showed that age,weight,BMI and treatment course were the suspected risk factors. The multivariate analysis results showed that age,BMI and treatment course were the risk factors of tacrolimus-related adverse reactions. Conclusion:The incidence of tacrolimus-related adverse reactions is relatively high,and if the patient is in advanced age,with poor nutritional status or long-term medication,the probability of adverse reactions will be increased.

Objective To offer a theory that supports the individualized tacrolimus dosage regimen by retrospectively investigating the influences of gene polymorphism and other clinical factors on tacrolimus concentration in renal transplant recipients. Methods A total of 280 renal transplant recipients were genotyped for CYP3A4?5, CYP3A4?6, CYP3A4?18B, CYP3A5?3, MDR1 1236C>T, MDR1 2677G>T/A, MDR1 3435C>T polymorphisms by PCR followed by restriction fragment length polymorphism (RFLP) analysis.PXR 6bp deletions (rs3842689) genotypes were determined by Allelic Special-Touch down PCR.Correlation between gene polymorphisms and tacrolimus concentrations was analyzed. Results The mutation frequency of CYP3A4?18B, CYP3A5?3, MDR1 1236C>T, 2677G>T/A, 3435C>T and PXR rs3842689 in the renal transplant recipients was 29.11%, 69.29%, 43.57%, 49.64%, 36.43% and 26.07%, respectively.Multiple regression analysis showed that, CYP3A5?3 and red blood cell count were associated with the value of C0/D of FK506, the best regression model was:D=C0/(-60.445 +95.777×CYP3A5 +34.938×RBC), and the equation could explain 38.8% of tacrolimus individual differences. Conclusion Gene polymorphism of CYP3A5?3 and red blood cell count may be responsible, in part, for the large interindividual variability of FK506 dose and concentration.

Objective To establish a sensitive method for the determination of rhodamine 123 (Rh123) in rat plasma by high performance liquid chromatography (HPLC).Methods The plasma samples were extracted by acetonitrile,and then separated on a Hypersil BDS C18 colunm (4.6 mm×100 mm,5 μm) equipped with a guard column kept at 25 ℃.The mobile phase consisted of acetonitrile and phosphate buffer (0.02 mol·L-1,pH4.0) (60:40) and was pumped at a constant rate of 1.0 mL·min-1.The peak was detected using a fluorescence detector set at FLD1A:Ex=485 nm,Em=546 nm.Results In this study,the method was validated for the Rh123 range of 0.1 to 32.0 μg·L-1,and the lower limit of quantitation (LLO Q) was 0.1 μg·L-1.The intra-and inter-day precisions for Rh123 were less than 7%,and the mean recoveries of Rh123 were 87.93%,89.03%,86.11% at low,mid,and high concentrations,respectively.Conclusion A simple,rapid and reproducible HPLC method was developed for the determination of Rh123 in rat plasma,which was an applicable method in modeling and description of the possible pharmacological interactions between the medicines and P-glycolprotein transporter.

Objective To examine the effect of berberine hydrochloride ( BER ) on the pharmacokinetic profiles of midazolam,a substrate of CYP3A,in rats. Methods The rats were intragastrically given different doses of BER (50,100, 200 mg?kg-1) or ketoconazole (75 mg?kg-1) for 10 days.Single-pass duodenum perfusion of 20 mg?kg-1 MDZ was performed and the inguinal artery was cannulated for blood sampling.Plasma concentrations of MDZ and 1'-OH-MDZ were analyzed by high performance liquid chromatography ( HPLC) with the CYP3A inhibitor ketoconazole serving as positive control. Results BER (50,100,200 mg?kg-1) and ketoconazole (75 mg?kg-1) could significantly increase the AUC(0-t),AUMC(0-t)and Cmax of MDZ in a dose-dependent manner ( P<0.05) ,and reduce the clearance rate ( CLz ) of MDA and its apparent volume of distribution in the body ( Vz ) ( P<0. 05). But they failed to dramatically affect the half-life ( t1/2z ) and the peak time ( tmax ) of MDZ. Additionally,BER ( 100,200 mg?kg-1 ) and ketoconazole ( 75 mg?kg-1 ) could significantly dose-dependently decrease the AUC(0-t),AUMC(0-t)and Cmaxof 1'-OH-MDZ,and profoundly increase the CLz,tmax and Vz of 1'-OH-MDZ (P<0.05),but they had no remarkable influences on the t1/2z.The ratio of AUC(1'-OH-MDZ)/AUC(MDZ) was decreased with the increase of BER concentration. Conclusion BER can inhibit the in vivo metabolism of MDZ in a dose-dependant manner, which is associated with the suppression of the activity of CYP3A.

Objectives: To evaluate mice as experimental animal for Chlamydia pneumoniae, a common cause of acute respiratory infections in human.　Methods: Intranasal inoculation of Icr mice with C. Pneumoniae induced a prolonged course of lung infection, as demonstrated by persistence of lung pathology(60 days).　Results: Icr mice were susceptible to C. pneumoniae. Lung pathology was characterized by patchy interstitial pneumonitis with predominately neutrophil leukocyte infiltration in the early(7 days) and lymphocytes infiltration in the later stages(14 days later) of infection.　Conclusions:Icr mice were susceptible to C. pneumoniae and the mouse model is useful for the investigation of the pathogenesis of C. pneumoniae infection.

Objective:To explore the clinical effect and safety of interventional treatment of autogenous arteriovenous fistula(AVF) stenosis.Methods:From July 2017 to September 2018, 96 patients with arteriovenous fistula stenosis and occlusion admitted to Handan First Hospital, Hebei Province were retrospectively analyzed.All of them were dialysis patients with chronic renal failure.All patients underwent percutaneous balloon angioplasty via the cephalic vein.The success rate of technique, clinical success rate, perioperative complications and follow-up were observed.Results:(1) Technical success rate and clinical success rate: 90 patients were treated with percutaneous transluminal angioplasty (PTA) via the cephalic vein, the other 3 patients were treated with interventional therapy via the brachial artery, and 3 patients underwent reconstruction of internal fistula.The technical success rate was 93.8% (90/96), and the clinical success rate was 89.6% (86/96). (2) Perioperative complications: thrombosis in 4 cases, vasospasm in 3 cases.There were no serious complications such as vascular rupture, aneurysm, vascular dissection, and no perioperative death.(3) The first stage patency rate was 100% (90/90), 74.4% (67/90), 62.2% (56/90) and 46.7% (42/90) in 3, 6, 12 and 18 months after operation.Conclusion:Venipuncture can be used as the first choice for AVF stenosis interventional therapy because of its advantages of small trauma, no serious complications, no need of long-term compression at the puncture point, immediate dialysis, and avoidance of local hematoma and other complications caused by artery puncture.

Objective:To compare the clinical effect and perioperative complications of the treatment of autogenous arteriovenous fistula stenosis by arterial and venous approach.Methods:The clinical data of 120 patients with AVF stenosis and occlusion who were treated with interventional therapy and met the inclusion criteria were collected and analyzed by retrospective case-control study.from September 2017 to August 2018, 60 patients with internal fistula stenosis were treated by transarterial approach (arterial approach group), and from September 2018 to may 2019, 60 patients were treated with a new surgical scheme(venous approach group). The operation success rate, perioperative complications and patency rate of 3, 6, 12 months after operation were compared between the two groups.Results:(1) The technical success rate was 96.7% (58/60) and the clinical success rate was 91.7% (55/60) in the arterial approach group, and 95.0%(57/60) and 93.3%(56/60) in the venous approach group.There was no significant difference in the technical success rate and clinical success rate between the two groups ( P=0.718 and 1.000, respectively) (2) Perioperative complications: in the arterial approach group, 3 patients had hematoma at the puncture point, 2 pseudoaneurysms and 5 thrombosis.There were 3 patients with thrombosis in the venous access group, and the difference in the incidence of complications between the two groups was statistically significant (χ 2=4.227, P=0.036). (3)The primary patency rates at 3, 6 and 12 months after operation were 95.0%(57/60), 75.0%(45/60) and 60.0%(36/60) in the arterial approach group, and 96.7%(58/60), 71.7%(43/60) and 61.7%(37/60) in the venous access group, respectively.There was no statistically significant difference between the two groups ( P=0.718, 0.749, 0.885). Conclusion:The interventional treatment for autogenous arteriovenous fistula stenosis through artery and vein approach can achieve good effect.There were many complications during the perioperative period, It is suggested that venous approach is preferred.

Objective To investigate the correlation between serum osteoblastin(OPN)and angiopoietin-like protein 8(ANGPTL8)levels and hepatic fibrosis(HF)after interventional therapy-transcatheter arterial chemoembolisation(TACE)in patients with primary hepatocellular carcinoma(PHC).Methods 166 patients with PHC admitted between March 2021 and June 2023 were selected and divided into 92 cases with HF(observation group)and 74 cases without HF(control group)according to whether or not HF occurred after interventional therapy;enzyme-linked immunosorbent assay(ELISA)was used to determine the serum OPN and ANGPTL8 levels and to analyse the predictive value of the OPN and ANGPTL8 levels on HF.Pearson correlation was used to analyze the correlation between OPN and ANGPTL8 levels and biochemical indexes.The factors influencing the occurrence of HF were analyzed by multi-factor Logistics regression.ROC curve was used to analyze the predictive value of OPN and ANGPTL8 for HF.Results Serum OPN[(74.56±11.56)ng/ml],ANGPTL[(42.78±5.23)ng/ml],ALT[(62.24±9.56)U/L],AST[(42.88±8.23)U/L],HA[(252.98±52.44)ng/L],LN[(152.64±26.45)ng/L],PC Ⅲ[(16.54±3.46)ng/L]and Ⅳ-C[(152.78±21.23)ng/L]in observation group were significantly higher than the control group[(57.89±9.68)ng/ml,(35.46±4.78)ng/ml,(49.46±7.46)U/L,(31.48±7.26)U/L,(192.56±23.88)ng/L,(124.48±11.23)ng/L,(11.26±2.23)ng/L and(126.45±18.56)ng/L].The differences between the two groups were statistically significant(P＜0.05).The AUC of serum OPN,ANGPTL8 and the combination of the two in predicting the occurrence of HF were 0.914,0.920 and 0.978,respectively,and the AUC of OPN combined with ANGPTL8 in predicting the occurrence of HF was higher than the AUC of the two separately(P＜0.05).Conclusion The levels of serum OPN and ANGPTL8 of patients with PHC are closely associated with the occurrence of HF,and the two are HF occurrence influencing factors and can be used as indicators to predict the occurrence of HF.

OBJECTIVETo evaluate mice as experimental animals for Chlamydia pneumoniae (C. pneumoniae) infection and investigate the pathogenesis of C. pneumoniae derived pneumonitis.

METHODSIcr mice were inoculated with the C. pneumoniae strain, CWL-029, either intranasally or intravenously. After a single dose inoculation, mice were killed on the 1st, 3rd, 7th, 14th, 21st, 28th and 60th days. The pathological changes in lung tissue were analyzed.

RESULTSThe Icr mice were shown to be susceptible to C. pneumoniae. Inoculation into mice with C. pneumoniae induced a prolonged course of lung infection, as demonstrated by persistence of lung pathology (up to 60 days). Via intranasal inoculation of mice, lung pathology was characterized by patchy interstitial pneumonitis with predominantly neutrophil leukocyte infiltration early (within the first 7 days) and lymphocyte infiltration in the later stages (14 days later) of infection. After intravenous inoculation, a similarly developed interstitial pneumonitis was observed, but it was milder and patchier, especially in early stages. C. pneumoniae DNA was detected by polymerase chain reaction (PCR) intermittently in the lung tissue. Inoculated mice developed serum IgG antibody responses.

CONCLUSIONThe Icr mice were susceptible to C. pneumoniae, resulting in a pulmonary infection characterized by interstitial pneumonitis, occurring most strongly via intranasal inoculation.

Animals ; Chlamydia Infections ; etiology ; pathology ; Chlamydophila pneumoniae ; DNA, Bacterial ; analysis ; Lung ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Pneumonia, Bacterial ; etiology ; pathology ; Polymerase Chain Reaction

Objective:To investigate the pathological characteristics and the clinical significance of novel coronavirus (2019-nCoV)-infected pneumonia (termed by WHO as coronavirus disease 2019, COVID-19).Methods:Minimally invasive autopsies from lung, heart, kidney, spleen, bone marrow, liver, pancreas, stomach, intestine, thyroid and skin were performed on three patients died of novel coronavirus pneumonia in Chongqing, China. Hematoxylin and eosin staining (HE), transmission electron microcopy, and histochemical staining were performed to investigate the pathological changes of indicated organs or tissues. Immunohistochemical staining was conducted to evaluate the infiltration of immune cells as well as the expression of 2019-nCoV proteins. Real time PCR was carried out to detect the RNA of 2019-nCoV.Results:Various damages were observed in the alveolar structure, with minor serous exudation and fibrin exudation. Hyaline membrane formation was observed in some alveoli. The infiltrated immune cells in alveoli were majorly macrophages and monocytes. Moderate multinucleated giant cells, minimal lymphocytes, eosinophils and neutrophils were also observed. Most of infiltrated lymphocytes were CD4-positive T cells. Significant proliferation of type Ⅱ alveolar epithelia and focal desquamation of alveolar epithelia were also indicated. The blood vessels of alveolar septum were congested, edematous and widened, with modest infiltration of monocytes and lymphocytes. Hyaline thrombi were found in a minority of microvessels. Focal hemorrhage in lung tissue, organization of exudates in some alveolar cavities, and pulmonary interstitial fibrosis were observed. Part of the bronchial epithelia were exfoliated. Coronavirus particles in bronchial mucosal epithelia and type Ⅱ alveolar epithelia were observed under electron microscope. Immunohistochemical staining showed that part of the alveolar epithelia and macrophages were positive for 2019-nCoV antigen. Real time PCR analyses identified positive signals for 2019-nCoV nucleic acid. Decreased numbers of lymphocyte, cell degeneration and necrosis were observed in spleen. Furthermore, degeneration and necrosis of parenchymal cells, formation of hyaline thrombus in small vessels, and pathological changes of chronic diseases were observed in other organs and tissues, while no evidence of coronavirus infection was observed in these organs.Conclusions:The lungs from novel coronavirus pneumonia patients manifest significant pathological lesions, including the alveolar exudative inflammation and interstitial inflammation, alveolar epithelium proliferation and hyaline membrane formation. While the 2019-nCoV is mainly distributed in lung, the infection also involves in the damages of heart, vessels, liver, kidney and other organs. Further studies are warranted to investigate the mechanism underlying pathological changes of this disease.