This paper is to report the development of a high-throughput in vitro system to screen hPXR/CAR mediated CYP2B6 drug inducers, and the application of it into the quick determination of induction activity toward CYP2B6 by various commonly used traditional Chinese medicines (TCMs) extract. Dual reporter gene assays were performed. The hPXR/CAR expression vectors and the reporter vector pGL3-CYP2B6-Luc involved in the distal and proximal promoters of CYP2B6 were co-transfected into HepG2 cells. Relative luciferase activities in cell lysate were analyzed after 48 h treatment of blank vehicle or drugs to determine the induction activity toward CYP2B6 by various commonly used TCMs extract. The positive hPXR/hCAR activators rifampicin and CITCO were applied to make sure that the reporter gene model was successfully established. Then 5 kinds of commonly used TCM extracts and 1 herbal compound were successfully investigated, some were found to activate hPXR or hCAR and therefore have the potential to induce CYP2B6 enzyme. This is the first domestic article to report the hCAR3-mediated CYP2B6 induction model and the establishment of a reporter gene system for hPXR/CAR-mediated CYP2B6 induction can be an effective and systemic in vitro method to investigate the drug inducers of CYP2B6 and to explain the mechanism involved.
Aryl Hydrocarbon Hydroxylases ; genetics ; metabolism ; Cytochrome P-450 CYP2B6 ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Genes, Reporter ; Genetic Vectors ; Hep G2 Cells ; High-Throughput Screening Assays ; Humans ; Luciferases ; genetics ; metabolism ; Oximes ; pharmacology ; Plants, Medicinal ; chemistry ; Plasmids ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; Receptors, Steroid ; genetics ; metabolism ; Rifampin ; pharmacology ; Thiazoles ; pharmacology ; Transfection

OBJECTIVETo evaluate the diagnosis and management of adrenal myelolipoma.

METHODSThe clinical data of 26 cases were analyzed retrospectively and the selected articles were reviewed. There were no specific clinical symptom and endocrine abnormality, except increasing catecholamine in 2 cases. All cases but two were diagnosed by B-model ultrasound scanning (B-US), CT or magnetic resonance imaging (MRI).

RESULTSTwenty-six cases were surgically treated, the diameter of the tumor was 5 - 10 cm, simple tumor resection was performed in 16 cases, and complete adrenal resection was performed in 10 cases. All the operated cases were proved by pathologists. The duration of follow-up was from 6 - 28 months after surgery. No recurrence was observed.

CONCLUSIONSThe diagnosis of adrenal myelolipoma could be established based on B-US, CT or MRI. The tumors whose diameter is larger than 5 cm in size should be removed.

Adrenal Gland Neoplasms ; diagnosis ; surgery ; Adrenalectomy ; Adult ; Female ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Myelolipoma ; diagnosis ; surgery ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo observe the changes of sex hormones and sexual function in male patients with Graves' disease (GD) after Radioiodine-131 (I-131) therapy.

METHODSThirty-four male GD patients, aged 21 -40 (32.3 +/- 6.7) years, were treated with I-131 at the dose of 111 - 407 (237.8 +/- 51.8) MBq. The levels of serum sex hormones were measured, and the patients'scores on erectile function (IIEF-5) were obtained before and 3 and 6 months after the treatment. Another 20 healthy men aged 25 - 37 (31 +/- 3.1) years were enlisted as controls.

RESULTSThe baseline levels of estrogen (E2), testosterone (T) and luteinizing hormone (LH) were (132.5 +/- 40.4) pmol/L, (21.6 +/- 4.6) nmol/L and (10.1 +/- 4.4) IU/L in the GD patients, significantly higher than (80.4 +/- 31.2) pmol/L, (14.5 +/- 4.2) nmol/L and (6.2 +/- 1.9) IU/L in the healthy controls (P < 0.05). The E2, T and LH levels showed a significant decrease in the GD patients after 3 months of treatment ([110.2 +/- 20.6] pmol/L, [17.7 +/- 5.5] nmol/L and (9.4 +/- 3.9) IU/L, P < 0.05), but exhibited no statistically significant differences from the healthy controls at 6 months ([82.6 +/- 30.1] pmol/L, [13.8 +/- 3.4 ] nmol/L and [6.6 +/- 1.5] IU/L, P > 0.05). The IIEF-5 score of the GD patients was 5 - 25 (15.5 +/- 3.5) before I-131 treatment, significantly lower than that of the controls (19 - 25, 24 +/- 0.5) (P < 0.05), and it was 8 - 25 (19.5 +/- 1.0) at 3 months and 10 - 25 (23.5 +/- 1.5) at 6 months, significantly higher in the latter than in the former (P < 0.05), and with no significant difference between the 6-month treated patients and the healthy controls (P > 0.05).

CONCLUSIONThe E2, T and LH levels are increased while the IIEF-5 score decreased markedly in male GD patients. Six-month treatment with I-131 can not only restore the E2, T and LH levels to normal but also significantly improve the patient's sexual function.

Adult ; Case-Control Studies ; Estrogens ; blood ; Follicle Stimulating Hormone ; blood ; Graves Disease ; blood ; therapy ; Humans ; Iodine Radioisotopes ; therapeutic use ; Luteinizing Hormone ; blood ; Male ; Testosterone ; blood ; Young Adult

Objective To analyze the magnetic resonance imaging (MRI) of the spinal cord and clinical characteristics in patients with autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy.Methods A total of 1 040 samples of cerebrospinal fluid (CSF) and sera collected in the Second Affiliated Hospital of Guangzhou Medical University from March 2013 to June 2018 were tested with tissue-and cell-based assays,and 42 patients were found positive for GFAP-IgG.The clinical data and MRI characteristics of the spinal cord of 19 patients who were positive for GFAP-IgG in CSF with autoimmune GFAP astrocytopathy and lesions in the spinal cord were retrospectively reviewed.Results There were 12 females and seven males among the 19 patients,with onset age of (44±17) years.The main manifestations of these patients included limb weakness (14/19),abnormal vision (5/19),headache (4/19),seizure (4/19),dementia (3/19),etc.On MRI of the spinal cord,five patients showed involvement in the cervical cord alone,eight showed involvement in the thoracic cord alone and six had both cervical and thoracic segment involvement.Fifteen patients had longitudinally extensive myelitic abnormalities (≥3 vertebral segments long).Seven enhancement patterns were encountered.Lesions were displayed in the spinal cord and brain in eight patients.Central gray matter involvement in the spinal cord was found in all the 19 patients.Conclusions Autoimmune GFAP astrocytopathy more frequently presents in females than in males.MRI of the spinal cord has complex presentations and longitudinally extensive myelitic abnormalities usually.Patients often show central gray matter involvement in the spinal cord.Myelitic abnormalities present more often in thoracic segment than in cervical segment.Abnormalities in lumbar segment are less encountered.

Objective To analyze the factors associated with pain after arthroscopic rotator cuff bridge suture.Methods According to the inclusion and exclusion criteria,the data of 112 patients with unilateral rotator cuff injury who received arthroscopic bridge suture in our department were collected and were investigated in the form of telephone follow-up.In this study,SPSS 23.0 was used to input data and conduct statistical analysis.Logistic regression analysis was used to analyze the correlation between the above influencing factors and postoperative pain.Results A total of 112 patients were included for statistical analysis,single factor analysis revealed,including course of disease,smoking history,preoperative University of California,Los Angeles(UCLA)score,Constant score,numeric rating scale(NRS),size of rotator cuff tear,whether it was full-thickness tear and degree of tendon retraction might be related to postoperative pain(P<0.05).The age,gender,body mass index(BMI),drinking history,diabetes and hypertension were not related to postoperative pain(P>0.05).Multiple linear regression analysis concluded that there were four factors related to postoperative pain,and the correlation degree was preoperative NRS,preoperative UCLA score,tear size and smoking history.Conclusion The causes of postoperative pain after arthroscopic rotator cauff repair are complex and diverse.Analyzing the cause of postoperative pain can effectively reduce the pain of patients and promote the recovery of shoulder joint function.

BACKGROUNDNurr1 plays an essential role in the development, survival, and function maintenance of midbrain dopaminergic (DA) neurons, and it is a potential target for Parkinson's disease (PD). Nurr1 mRNA can be detected in peripheral blood mononuclear cells (PBMCs), but whether there is any association of altered Nurr1 expression in PBMC with the disease and DA drug treatments remains elusive. This study aimed to measure the Nurr1 mRNA level in PBMC and evaluate the effect of Nurr1 expression by DA agents in vivo and in vitro.

METHODSThe mRNA levels of Nurr1 in PBMC of four subgroups of 362 PD patients and 193 healthy controls (HCs) using real-time polymerase chain reaction were measured. The nonparametric Mann-Whitney U-test and Kruskal-Wallis test were performed to evaluate the differences between PD and HC, as well as the subgroups of PD. Multivariate linear regression analysis was used to evaluate the independent association of Nurr1 expression with Hoehn and Yahr scale, age, and drug treatments. Besides, the Nurr1 expression in cultured PBMC was measured to determine whether DA agonist pramipexole affects its mRNA level.

RESULTSThe relative Nurr1 mRNA levels in DA agonists treated subgroup were significant higher than those in recent-onset cases without any anti-PD treatments (de novo) (P < 0.001) and HC groups (P < 0.010), respectively. Furthermore, the increase in Nurr1 mRNA expression was seen in DA agonist and L-dopa group. Multivariate linear regression showed DA agonists, L-dopa, and DA agonists were independent predictors correlated with Nurr1 mRNA expression level in PBMC. In vitro, in the cultured PBMC treated with 10 μmol/L pramipexole, the Nurr1 mRNA levels were significantly increased by 99.61%, 71.75%, 73.16% in 2, 4, and 8 h, respectively (P < 0.001).

CONCLUSIONSDA agonists can induce Nurr1 expression in PBMC, and such effect may contribute to DA agonists-mediated neuroprotection on DA neurons.

Adult ; Aged ; Aged, 80 and over ; Dopamine Agonists ; therapeutic use ; Female ; Humans ; Leukocytes, Mononuclear ; drug effects ; metabolism ; Male ; Middle Aged ; Nuclear Receptor Subfamily 4, Group A, Member 2 ; genetics ; Parkinson Disease ; drug therapy ; genetics ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVETo investigate the mechanism underlying myofibroblast differentiation induced by transforming growth factor (TGF) beta1 in obliterative bronchiolitis following lung transplantation.

METHODSHeterotopic tracheal transplantation was performed in Smad3 wild-type and knock-out mice to simulate the lung transplantation in human. Murine tracheal fibroblasts cultivated in primary culture were used for in vitro study. Immunohistochemistry, immunocytochemistry, Western Blotting, RT-PCR and DNA electrophoresis mobility gel shift assay were conducted to detect the expression of alpha-smooth muscle actin (alphaSMA), the marker of fibroblast-myofibroblast differentiation, and the activation of Smad3, p38 and ERK1/2.

RESULTSIn affected airways of experimental obliterative bronchiolitis, abundant expression of alphaSMA were found. In vitro study for tracheal fibroblasts, the activation of Smad3 by TGF-beta1 presents as three major forms, phosphorylation, nuclear translocation and DNA binding. In Smad3 wild-type fibroblasts, TGF-beta1 induces the increase of the myofibroblasts transformation, characterized by the elevation of alphaSMA, both at transcription and protein level. While in Smad3 knock-out fibroblasts, the transformation of myofibroblasts induced by TGF-beta1 is significantly decreased (t = 2.080, P = 0.027; t = 1.982, P = 0.032), but not completely abolished. Further study in Smad3-deficient fibroblasts demonstrates that p38 and ERK1/2 could be activated by TGF-beta1 and result in fibroblast differentiation.

CONCLUSIONSTGF-beta1 could promote the transformation of fibroblasts into myofibroblasts in Smad3 dependent and independent signal pathways, especially the Smad3 dependent path, and result in the development of obliterative bronchiolitis.

Actins ; genetics ; metabolism ; Animals ; Blotting, Western ; Bronchiolitis Obliterans ; genetics ; pathology ; surgery ; Cell Differentiation ; drug effects ; Cells, Cultured ; Disease Models, Animal ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Immunohistochemistry ; Mice ; Mice, Inbred BALB C ; Mice, Knockout ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Smad3 Protein ; genetics ; metabolism ; Trachea ; cytology ; transplantation ; Transforming Growth Factor beta1 ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective Our study aimed to delineate the clinical and radiological features of patients with anti-myelin oligodendrocyte glycoprotein antibody (MOG-Ab)positive neuromyelitis optica spectrum disorder (NMOSD). Methods Fifty-seven patients with NMOSD and 29 patients with multiple sclerosis (MS) were collected. Data on clinical and radiological features of MOG-Ab positive patients with were analyzed retrospectively. Results MOG-Abs were present in 9/57 (15.8%) NMOSD patients and 2/29 (6.9%) MS patients. Both MOG and aquaporin-4 (AQP4) antibodies were positive in one case of NMOSD. There was no significant difference between the two groups (P＞0.05). There were more females than males having MOG-Ab positive NMOSD (females: males=7:1) and the average onset age was 41.4 ± 11.5 years. There was no significant difference in gender and age between MOG-Ab negative and AQP4-Ab positive groups(P＞0.05). The durations of disease were significantly shorter in either MOG-Ab positive NMOSD patients or MOG-Ab negative NMOSD patients than in AQP4-Ab positive group (P<0.05). Recurrence was the main disease pattern of all three groups and the frequency of recurrence was not significant different among three groups (P＞0.05). The incidence of optic neuritis was 62.5% in NMOSD patients with MOG-Ab positive and 43.5% in AQP4-Ab positive NMOSD patients (P＞0.05). There was no significant difference in the morphology and location of brain lesions among the three groups (P＞0.05). MOG-Ab positive NMOSD patients had long segment spinal cord lesions. The median length of the spinal cord lesions in the MOG-Ab positive group was similar to the other two groups (P＞0.05). Conclusions MOG-Ab positive NMOSD patients have higher proportion of females with shorter recurrence course, more likely complicated with optic neuritis. And the radiological features of brain and spinal cord were not specific to patients with AQP4-Ab positive.

Objective Surgical treatment of the urethral diverticulum is difficult and may be followed by serious postoperative complications such as urethro-vaginal fistula. The purpose of this study is to investigate the treatment of female urethral diverticulum and its clinical effect.Methods We retrospectively analyzed the clinical data on 20 cases of female urethral diverticulum confirmed and treated in our hospital from January 2010 to December 2017. We obtained the Overactive Bladder Symptom Score (OABSS) from the patients after admission, treated them by transvaginal urethral diverticulectomy under continuous epidural anesthesia, and compared the signs and symptoms before and after operation.Results All the patients showed significantly decreased incidences of perineal discomfort, frequent micturition, urgent urination, and dysuria after operation (P<0.05). The average OABSS was remarkably lower postoperatively than the baseline (4.9±1.8 vs 8.1±2.3, P<0.05). No such postoperative complications as hematuria, urinary fistula, urethral stricture and urinary incontinence were observed in any of the patients.Conclusion Transvaginal urethral diverticulectomy is safe and effective for the treatment of female urethral diverticulum.

Objective: To observe the clinical efficacy of Wumeiwan combined with Bazhentang in the treatment of obesity type 2 diabetes with Qi and Yin deficiencies, phlegm and stasis. Method: Totally 60 patients with type 2 diabetes mellitus were randomly divided into observation group and control group, with 30 cases in each group. Observation group was given Wumeiwan combined with modified Bazhentang in addition to Western medicine (metformin hydrochloride). The control group was treated with traditional Western medicine (metformin hydrochloride). The course of treatment is eight weeks. Fasting blood sugar (FPG),2 hPG (2 hPG),glycosylated hemoglobin (HbA1c),total cholesterol (TC),triglycerin (TG),body mass index (BMI),safety indicators (three major routine,liver and kidney functions) and clinical symptoms before and after treatment between two groups were compared. The clinical efficacy of two groups was evaluated. Result: The observation group had an effective rate of 93.3% (28/30),which was significantly higher than 73.3% (22/30) of the control group,with statistically significant differences (P<0.05). Compared with before treatment,the levels of FPG,2 hPG and HbA1c were significantly lower in two groups after treatment, and the treatment group was lower than control group (P<0.05). The levels of TC,TG,BMI and TCM syndromes were significantly reduced after treatment in two groups (P<0.05),and the levels of TC,TG,BMI and TCM syndromes were significantly lower than those in control group. The differences were statistically significant (P<0.05). Conclusion: Modified Wumeiwan combined with Bazhentang is safe and effective in treating patients with type 2 diabetes mellitus with Qi and Yin deficiencies, phlegm and stasis,and can reduce blood sugar,blood lipid,BMI level and relieve clinical symptoms of patients, and so worth promotion.