Studies on the inhibitory effect of isavuconazole on flumatinib metabolism in vitro and in vivo.

As the validated agent for the treatment of chronic myelogenous leukemia (CML), flumatinib is a novel oral tyrosine kinase inhibitor (TKI) with higher potency and selectivity for BCR-ABL1 kinase compared to imatinib. Many patients experience aspergillosis infection and they may start using isavuconazole, which is an inhibitor of CYP3A4. However, there is no study on their interaction in vitro and in vivo. In the present study, the concentrations of flumatinib and its major metabolite M1 were rapidly determined using an stable ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method. The half-maximal inhibitory concentration (IC50) was 6.66 µM in human liver microsomes (HLM), while 0.62 µM in rat liver microsomes (RLM) and 2.90 µM in recombinant human CYP3A4 (rCYP3A4). Furthermore, the mechanisms of inhibition of flumatinib in human liver microsomes, rat liver microsomes and rCYP3A4 by isavuconazole were mixed. Moreover, ketoconazole, posaconazole, and isavuconazole showed more potent inhibitory effects than itraconazole, fluconazole, and voriconazole on HLM-mediated flumatinib metabolism. In pharmacokinetic experiments of rats, it was observed that isavuconazole could greatly change the pharmacokinetic parameters of flumatinib, including AUC(0-t), AUC(0-∞), Cmax and CLz/F, but had no effect on the metabolism of M1. According to the results of in vitro and in vivo studies, the metabolism of flumatinib was inhibited by isavuconazole, suggesting that isavuconazole may raise the plasma concentration of flumatinib. Thus, it is important to take special care of the interactions between flumatinib and isavuconazole in clinical applications.

Evaluation of the inhibitory effect of quercetin on the pharmacokinetics of tucatinib in rats by a novel UPLC-MS/MS assay.

CONTEXT: Tucatinib (CYP2C8 substrate) and quercetin (CYP2C8 inhibitor) are two common drugs for the treatment of cancer. However, the effect of quercetin on the metabolism of tucatinib remains unknown. OBJECTIVE: We validated a sensitive method to quantify tucatinib levels in rat plasma based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), which was successfully employed to explore the effect of quercetin on tucatinib pharmacokinetics in rats. MATERIALS AND METHODS: An Acquity UPLC BEH C18 column was applied to achieve the separation of tucatinib and internal standard (IS) talazoparib after protein precipitation with acetonitrile. Then, we used this assay to investigate the effect of different doses of quercetin (25, 50 and 100 mg/kg) on the exposure of orally administered tucatinib (30 mg/kg) in 24 Sprague-Dawley (SD) rats, which were randomly divided into three quercetin pre-treated groups and one control group (n = 6). RESULTS: Our developed assay was verified in all aspects of bioanalytical method validation, involving lower limit of quantification (LLOQ), selectivity, accuracy and precision, calibration curve, extraction recovery, matrix effect and stability. After pre-treatment with 100 mg/kg quercetin, AUC0→t, AUC0→∞ and Cmax of tucatinib were remarkably increased by 75.4%, 75.8% and 59.1% (p < 0.05), respectively, while CLz/F was decreased significantly by 47.3% (p < 0.05) when compared with oral administration of 30 mg/kg tucatinib alone. This change is dose-dependent. CONCLUSIONS: This study will help better understand the pharmacokinetic properties of tucatinib with concurrent use with quercetin, and more clinical verifications were inspired to confirm whether this interaction has clinical significance in humans.

Nerve Growth Factor Protects Retinal Ganglion Cells Related to Inhibiting Endoplasmic Reticulum Stress by Inhibiting IRE1-JNK-CHOP Signaling Pathway.

BACKGROUND: Under various physiological conditions, endoplasmic reticulum stress can induce apoptotic cell death, leading to brain and retinal neuronal cell death, but the relations of ER stress-induced apoptosis and the nerve growth factor's therapeutic effect in Glaucoma optic neuropathy still unclear. METHODS: An endoplasmic reticulum stress model was established in ganglion cells using TG, the endoplasmic reticulum stress inducer. MTT assay and flow cytometry were used to detect the protective effect of NGF on retinal ganglion cells. Western blot was used to detect apoptosis-related proteins Bcl-2, Bad and endoplasmic reticulum stress-related proteins GRP78, IRE1, JNK and CHOP. RESULTS: MTT assay and flow cytometry showed NGF can protect the apoptosis of ganglion cells. Western blot analysis showed the level of pro-apoptotic protein Bad was decreased and anti-apoptotic protein Bcl-2 was increased after NGF treatment. Endoplasmic reticulum stress-induced proteins GRP78, IRE1, JNK and CHOP are counter- acted by NGF. CONCLUSION: NGF protects retinal ganglion cells related to inhibiting endoplasmic reticulum stress by inhibiting IRE1-JNK-CHOP signaling pathway.

Photodynamic Treatment of Colorectal Cancer Using Chlorin e6-Loaded Poly(lactide-co-glycolide)- Based Nanoparticles.

We prepared poly(lactide-co-glycolide) (PLGA) encapsulated with chlorin e6 (Ce6) in an effort to increase the stability and efficiency of photosensitizers for photodynamic therapy (PDT). We determined that Ce6-loaded PLGA nanoparticles (PLGA-Ce6 NPs) had drug-loading efficiency of 5%. The efficiency of encapsulation was 82%, the zeta potential was- 25 mV, and the average diameter was 130 nm. The encapsulation of Ce6 in PLGA nanoparticles showed excellent stability. The nanoparticles exhibited sustained Ce6 release profiles with 50% released at the end of 3 days, whereas free Ce6 showed rapid release within 1 day. Ce6 release patterns were controlled by encapsulation into PLGA. The uptake of PLGA-Ce6 NPs was significantly enhanced by endocytosis in the first 8 hours in the HCT-116 cell line. An intracellular reactive oxygen species assay revealed the enhanced uptake of the nanoparticles. An in vitro anti-tumor activity assay showed that the PLGA-Ce6 NPs exhibited enhanced phototoxicity toward HCT-116 cells and a slightly lower IC50 value in HCT-116 cells than Ce6 solution alone. Exposure of HCT-116 cell spheroids to PLGA-Ce6 NPs penetrated more profoundly and had better phototoxicity than pure drugs. These findings suggest that PLGA-Ce6 NPs might serve as PDT for colorectal cancer.

Therapeutic effects of Houttuynia eye drops combined with olopatadine hydrochloride eye drops on vernal keratoconjunctivitis.

Therapeutic effects and safety of houttuynia eye drops combined with olopatadine hydrochloride eye drops on vernal keratoconjunctivitis (VK) were evaluated. A total of 926 patients with VK were collected in the Eye Hospital of Wenzhou Medical University from July 2011 to January 2017. Patients were divided into group A, B and C according to the use of different eye drops. Group A included 276 patients who were treated with houttuynia eye drops; group B included 305 patients who were treated with olopatadine hydrochloride eye drops; group C included 345 patients who were treated with houttuynia eye drops and olopatadine hydrochloride eye drops. Treatment was performed for 14 days. Eye symptoms before and after treatment, and at 1 h, 7 days and 14 days after drug administration were compared among groups to evaluate the therapeutic effect. At 1 h after drug administration, highest excellent rate was observed in group C, followed by group A, and good outcome rate was significantly higher in group C than in the other two groups (P<0.05), and effective rate was also significantly higher in group C than in the other two groups (P<0.05). At 7 days after treatment, excellent rate was significantly higher in group C than in the other two groups (P<0.05), and effective rate was higher in group C than in group B (P<0.05). At 14 days after treatment, excellent rate was significantly higher in group C than in the other two groups (P<0.05), while lowest good outcome rate and ineffective rate were found in group C, and no significant differences were found between group A and B (P>0.05). Houttuynia ophthalmic solution and olopatadine hydrochloride eye drops can both achieve good effect in the treatment of VK, and combined use of them can increase the efficacy and shorten treatment period. So, the combined treatment should be popularized.

Molecular mechanism of the role of carbamyl erythropoietin in treating diabetic retinopathy rats.

The aim of the present study was to investigate the therapeutic effects of carbamyl erythropoietin (CEPO) and safflor yellow (SY) in the treatment of rats with diabetic retinopathy (DR) as well as exploring the mechanism of action. Male SD rats were used to establish a diabetes model and streptozotocin-induced retinopathy was also performed in rats. A total of 126 rats with DR were obtained, and model rats were randomly divided into the model (n=42), experimental (n=42) and control (n=42) groups. The rats in the model group were injected with saline, the rats in the experimental group were treated with CEPO, and the rats in the control group were treated with SY. After treatment for 2 weeks, the retinas were harvested for quantitative analysis of the mRNA expression levels of angiogenesis-promoting and -inhibiting molecules, apoptosis-promoting and -inhibiting molecules, and oxidative stress pathway-related factors by Reverse transcription-quantitative PCR (RT-qPCR). No significant differences in expression levels of hypoxia-inducible factor-1α (HIF-1α), vascular endothelial growth factor (VEGF), angiopoietin (Ang-1), tissue kallikrein (TKLK) and pigment epithelium-derived factor (PEDF) were observed between the experimental and model groups (P>0.05). The expression levels of apoptosis-promoting molecules Bcl-2 related X protein (Bax) and cysteine aspartate specific protease (caspase-3) mRNA in the retina of the experimental group was significantly lower than those in the control group (P<0.05). The expression levels of Bcl-2 and survivin mRNA were significantly higher in the experimental group than in the control group (P<0.05). The expression levels of the oxidative stress pathway nuclear factor erythroid 2 (NFE2)-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and NAD(P)H quinone dehydrogenase 1 (NQO1) mRNA were significantly higher in the experimental group than in the control group. Therefore, the therapeutic effects of CEPO in treating DR are better than those of SY. As a result, CEPO may inhibit apoptosis and oxidative stress damage of retinal tissue cells in DR rats without affecting angiogenesis.

Effects of a novel curcumin derivative on the functions of kidney in streptozotocin-induced type 2 diabetic rats.

OBJECTIVE: B6, an analog of curcumin, is a compound isolated from a traditional Chinese medicine Turmeric. In this paper, we aimed to explore the efficacy of B6 on diabetic nephropathy and the related mechanisms. MATERIALS AND METHODS: The effects of B6 were studied on fast-blood glucose, serum creatinine, urea nitrogen, urine albumen/24 h, pathological changes of main organs, the levels of ACE2 and ACE2 mRNA in the rat model of diabetes induced by streptozotocin. RESULTS: The results showed that B6 treatment could reduce serum creatinine, urea nitrogen, urine albumen/24 h, decrease the level of AngII, improve the renal pathological changes in diabetic rats and increase the levels of ACE2 and ACE2 mRNA. CONCLUSION: These results suggested B6 could protect the renal function of diabetic rats. This study provided scientific basis for the further researches and clinical applications of B6.

Effects of puerarin on the retina and STAT3 expression in diabetic rats.

The effects of puerarin on electroretinogram, oxidative stress and STAT3 expression were determined, in diabetic rat retina and serum. Forty Sprague-Dawley rats were randomly divided into the normal control (NC), the diabetic model (DM), the low dose (250 mg/kg) puerarin (LP) or the high dose (500 mg/kg) puerarin group (HP). A diabetic rat model was induced by streptozotocin and animals were continuously treated for 4 weeks; fasting blood glucose was measured at 2 and 4 weeks after modeling. An electroretinogram and serum and tissue levels of glucose, insulin, superoxide dismutase (SOD), malondialdehyde (MDA) and total antioxidant capacity (T-AOC) were measured; real-time PCR and ELISA were used to determine STAT3 mRNA and protein expression, respectively, from the retina. The blood glucose and insulin levels in the puerarin groups were significantly lower and higher, respectively than that in the DM group. The amplitude of b-wave of electroretinogram in the DM and the LP groups was significantly lower than that in the NC group; in the LP and HP groups it was significantly higher than the DM group. The serum and retinal tissue activity of SOD and MDA was significantly lower and higher, respectively, in the DM group compared to the NC group; both the LP and HP groups had significantly higher SOD and lower MDA than the DM group. The levels of STAT3 mRNA and protein levels in the DM, LP and HP groups were significantly higher than the NC group; and levels of STAT3 mRNA and protein expression were significantly lower in the LP and HP groups than the DM group. In summary, puerarin can reduce the oxidative stress damage of the retina, and its mechanism is related to the inhibition of STAT3 expression.

[Research on the patterns of upper airway obstructive levels by drug-induced sleep endoscopy].

OBJECTIVE: To identify the patterns of airway collapse in patients with obstructive sleep apnea hypopnea syndrome(OSAHS) by dexmedetomidine induced sleep endoscopy. METHODS: Forty-five obstructive sleep apnea patients diagnosed by polysomnography were given dexmedetomidine intravenously. Once the patient was sedated in dorsal position, the electronic nasopharyngoscope was inserted transnasally and positioned on five levels of the upper airway sequentially (velum, oropharyngeal lateral wall, tongue base, epiglottis and larynx) to observe and document the collapse. Each level should be observed no less than three apneas. The degree of airway narrowing was calculated by using the ImageTool. No obstruction was defined when the degree of airway narrowing <50%, and complete obstruction when ≥ 75%. RESULTS: In 45 patients with OSAHS, 1 case showed no obstruction on any level, 6 cases demonstrated obstructions on single level only, and 38 cases demonstrated complete obstructions on multilevel, including 17 cases with complete obstructions on two levels, 15 cases complete obstructions on three levels, and 6 cases complete obstructions on four levels. The patterns of collapse found in the trial were: (1) circumferential stricture by velum collapse was found in 43 patients, and 41 cases showed complete obstructions; (2) the side wall of oropharynx all collapsed in a lateral configuration, and 32 cases showed complete obstructions on this level; (3) anteroposterior swallowing tongue base was common, 11 cases showed partial obstructions on level of tongue base, and 10 cases complete; (4) epiglottic collapses occurred in lateral configuration folding as V shape; in anteroposterior configuration, epiglottis met posterior wall of the pharynx due to swallowing tongue base; the server soften epiglottis obstructed the entrance of the larynx, while the mild soften epiglottis and the collapsed side wall of pharynx came into being obstructions in concentric configuration; (5) the arytenoid area and aryepiglottic fold mucosa inwardly covered the glottis when the obstruction occurred in the larynx. CONCLUSIONS: The patterns of hypopharynx obstructions in OSAHS patients are multifarious. Lateral oropharyngeal wall, epiglottic and tone base collapse play an important role in the obstructions. The laryngeal obstruction can also be observed.

[The expression and clinical pathological significance of PDCD in laryngocarcinoma].

OBJECTIVE: To investigate the expression and clinical pathological significance of PDCD4 in laryngocarcinoma tissue and its potential significance to clinic. METHOD: Western-blotting and immunohistochemistry ana lyse to measure the protein expression of PDCD4 in 54 cases of laryngocarcinoma tissues (studying group) and their paraneoplastic normal tissues (control group). The correlations of PDCD4 with clinical pathological parameters were analyzed. RESULT: PDCD4 protein was positively expressed in paraneoplastic normal tissue while which was lost or decreased in laryngocarcinoma tissue by Western blot and immunohistochemistry. Immunohistochemistry assay showed the location of PDCD4 protein in cells was different between the studying group and the control group. The expression level of PDCD4 was related to the pathological grades of the laryngocarcinoma. It's higher in the well-differentiated tumor group than that in the poorly differentiated ones. But the expressions of PDCD4 were no differences among other clinical parameters including sex, age, clinical classification, clinical stage and the cervical lymphonodus who had been metastases or not. CONCLUSION: PDCD4 gene is anti-oncogene. It may play an important role in the pathogenesis and development of laryngeal carcinoma and it may be a new target of therapy for laryngo carcinoma.

Comparison of the retinal modulation transfer function between amblyopes successfully corrected and normal subjects.

AIM: To analyze the retinal modulation transfer function between amblyopes whose visual acuity was corrected to 5.0 and normal subjects at the same age. METHODS: RM-800 used to detect contrast sensitivity was adopted to measure MTF of 96 amblyopes (96 eyes) whose visual acuity was corrected to 5.0 and 80 normal controls (80 eyes) at the same age under six interference fringes (IVA=0.06, 0.1, 0.2, 0.4, 0.6, 0.8). RESULTS: The functional values of amblyopes were significantly lower than those of normal subjects in every fringe (P<0.01), especially in medium and high frequency. CONCLUSION: For amblyopes, MTF was still abnormal after stopping the treatments.

Dynamic monitoring of total plasma homocysteine in spontaneously hypertensive rats by LC-MS.

Hypertension has been recognized to be closely related to plasma homocysteine levels (tHcy). Spontaneously hypertensive rats (SHR) are used widely for hypertension research, but it is unclear whether hypertension is related to high levels of tHcy in rat plasma. To test whether hyperhomocysteinemia occurs in SHR we dynamically measured plasma total homocysteine (tHcy) in SHR by liquid chromatography-tandem mass spectrometry (LC-MS). This analytical method has good linearity within the range of 1-100 micromol/L for tHcy in rat plasma with a correlation coefficient of R = 0.9975. After dynamic monitoring (12 weeks) on the plasma tHcy in SHR and Wistar-Kyoto rats, we found that there was no significant difference in tHcy level between SHR and Wistar-Kyoto rats, which was 6.98 +/- 1.82 micromol/L and 8.04 +/- 1.64 micromol/L, respectively. And there was no significantly high level of plasma tHcy in SHR.

[Substance P and its receptors are involved in the effect of histamine H3 receptor agonist, IMETIT on nasal allergic symptoms in guinea pigs].

OBJECTIVE: To explore the influence of histamine H3 receptor agonist, IMETIT and simultaneous use of IMETIT and H1-receptor antagonist, Loratadine, on the symptoms of allergic rhinitis (AR) and substance P(SP) secretion and expression of SP receptor (SP-R) mRNA in AR model in guinea pigs. METHODS: Guinea pigs were divided randomly into 4 groups: AR group (group A), IMETIT group (group B), Loratadine group (group C) and IMETIT+Loratadine group (group D). The severity of AR was assessed by determining the extent of three markers of allergic symptoms (sneezing, nasal rubbing and nose blocking). The changes in the nasal mucosa were studied by pathological methods. The expression of positive cell of SP was detected by immunohistochemistry. SP-R mRNA expression in nasal mucosa was used to do reverse transcriptive-polymerase chain reaction (RT-PCR). Statistical analysis was performed using a SPSS 13.0 software. RESULTS: In Group B, the mean (x ± s) number of sneeze [(15.0 ± 1.3) times], scratching nose [(16.5 ± 2.3) times] and respiratory frequency [(76.3 ± 4.1) times/min] were significantly improved than those in group A [(23.5 ± 2.6) times, (26.1 ± 4.1) times and (66.5 ± 5.8) times/min, respectively), P value were 0.000, 0.000 and 0.001, respectively]. The numbers of SP-positive cells [(11.6 ± 3.6)/HP] and SP-R mRNA expression (0.64 ± 0.04) in group B were reduced significantly compared to group A [(27.1 ± 9.7)/HP, (0.83 ± 0.03), P value were 0.000, 0.000, respectively]. Sneeze [(10.0 ± 2.3) times], scratching nose [(11.8 ± 1.7) times] and respiration [(90.0 ± 5.0) times/min] in Group D were improved significantly than those in group B (P value were 0.000, 0.002 and 0.000, respectively). SP-positive cells [(2.0 ± 1.7)/HP] and SP-R mRNA expression (0.52 ± 0.06) in Group D compared with group B were also significantly reduced (P value were 0.012 and 0.000, respectively). Pathological changes in guinea pig nasal mucosa in group B, group D were alleviated than those in group A. The combination of IMETIT and Loratadine had a synergistic effect on these effects (F value were 11.59, 8.28, 5.61, 5.48, 6.50, respectively, P value were 0.002, 0.008, 0.025, 0.027, 0.017). CONCLUSIONS: IMETIT and the combination of IMETIT with Loratadine can effectively relieve the symptoms of AR in guinea pigs, its mechanism may be relevant to reduce SP secretion and the expression of SP-R mRNA, and the two has a synergistic effect. It may be useful as a novel therapeutic approach in nasal allergy.

[Effect of H3R agonist, IMETIT on allergic rhinitis in guinea pigs].

OBJECTIVE: To discuss the treatment of H3R agonist, IMETIT, on the allergic rhinitis(AR) ,and the influence to mRNA of Substance P(SP) and Substance P Receptor (SP-R) in AR model of guinea pigs. METHOD: The severity of AR was assessed by allergic symptoms (sneezing, nasal rubbing and nose blocking). The changes in the nasal mucosa were studied by pathological methods. The expression of SP positive cell was detected by immunohistochemistry, and the expression of SP-R mRNA was detected by reverse transcriptive polymerase chain reaction (RT-PCR). RESULT: Histamine H3R agonists, IMETIT can effectively improve the AR symptoms, sneezing, nasal itching, nasal congestion, reduce the pathological changes in the nasal mucosa, cut down the SP secretion and SP-R mRNA expression. CONCLUSION: Histamine H3R agonist, IMETIT can effectively relieve the symptoms of AR in guinea pigs, which is related to reducing SP secretion and SP-R mRNA expression.

[Expression of tenascin in human nasal polyp tissue and primary studies of its clinical significance].

OBJECTIVE: To explore the pathogenesis of nasal polyposis and the expression of tenascin in human nasal polyps. METHOD: Tenascin in nasal polyp tissues from twenty-four polyposis patients and tenascin in inferior turbinates tissues from fifteen chronic hypertrophic rhinitis patients were detected with immunohistochemistry (sp) method. The inferior turbinate mucosa of five healthy volunteers were selected as control. RESULT: Tenascin were expressed in the mucosa epithelial cells and serous cells from nasal polyps and inferior turbinates of chronic hypertrophic rhinitis; The level of tenascin in the mucosa epithelial cells from nasal polyps were significantly(P < 0.01); The level of tenascin in the serous cells from nasal polyps were significantly higher than that in inferior turbinates of chronic hypertrophic rhinitis(P < 0.01); Tenascin were hardly detected in nasal mucosa from healthy volunteers; The tenascin positivity in glandular cell was significantly higher in nasal polyps than in inferior turbinates of chronic hypertrophic rhinitis (P < 0.05). CONCLUSION: There is close relationship between tenascin and nasal polyps. Tenascin is strongly expressed in nasal polyps and its mucosa, where it could be produced by mucosa epithelial cells and serous cells.

[Surgery of infratemporal fossa tumors].

OBJECTIVE: To investigate the effects of surgical approach on tumor with advanced infratemporal fossa. METHOD: Four different kinds of surgical approaches were selected and used according to preoperative clinical and radiographic findings and its invading structures. RESULT: The survival rate according to Kaplan-meire showed that the two years survival rate for benign tumor were 100%, but malignant tumor were 47.1%. CONCLUSION: Surgery of the infratemporal fossa tumors appears to be safe and complete removal of the tumor invading lateral skull bone and its surrounding structures is necessary. The adjuvant chemotherapy and radiotherapy are worth while procedure for postoperative of malignant tumors.