The dynamic level of immunosuppressants such as alpha-fetoprotein(α-FP), gestation associated protein, progesterone, estradiol, chorionic gonadotropin, etc. of peripheral sera and the regulatory mechanism of cellular immunity in the peripartum stages in sows were determined by advanced radioimmunoassay (RIA) and immunochemistry methods in this study. The effects on lymphocyte cultivated in vitro by the immunosuppressants mentioned above were tested by cell culture technology, respectively. The results were as follows: (1) α-FP: The level of α-FP of peripheral sera in the stage of ante-parturition was slightly higher than that of post-parturition in sows, but it didn't show statistical difference. The concentration of α-FP of peripheral sera in conceptus sows had no obvious effect on lymphocyte cultivated in vitro, and it indicated that α-FP was not the major immunosuppressant in the pregnancy of sows. However, the α-FP, whose concentration was equal to that of amniotic fluid, had obvious inhibition on lymphocyte cultivated in vitro. (2) Gestation associated protein: The level of gestation associated protein was maintained highly (100-162 mg/L) in peripheral sera and amniotic fluid in the stage of ante-parturition in sows, then descended gradually after parturition and disappeared step by step 5-10 d after parturition. The gestation associated protein, whose concentration were equal to that of peripheral sera and amniotic fluid, were added to lymphocyte culture in vitro, and the effect on lymphocyte activity was not observed yet. This showed that gestation associated protein was not the major immunosuppressant in the peripartum stage in sows. (3) Estradiol and progesterone: Estradiol or progesterone at the level of sera in pregnant sows, could lower the rate of lymphocyte transformation and the level of cellular immunity. (4) Chorionic gonadotropin: The level of chorionic gonadotropin of peripheral sera in the stage of ante-parturition was slightly higher than that of post-parturition in sows, but it didn't have statistical difference. The chorionic gonadotropin, whose concentration was equal to that of sera, was not yet observed obvious effect on lymphocyte cultivated in vitro. (5) Added the five above-mentioned immunosuppressants to lymphocyte culture in vitro, whose concentration were equal to those of sera in pregnant sows, they could drop the rate of lymphocyte resette formation and lymphocyte transformation, lower the level of cellular immunity and reduce the concentration of IL-2 which synthesized and secreted by lymphocyte in culture remarkably. Their inhibitory intensity of cellular immunity was evidently higher than that of any single immunosuppressant.

To investigate the immunological enhancement effects of IL-2 with different doses in chicken Shigella vaccine on the duodenum mucosa in Gushi chicken,parental line Gushi chicken were grouped and inoculated with Shigella vaccines including different dose of IL-2.And then the distribution and change of SIgA positive cells and its secretion in the duodenum of the chicken were observed with the immunohistochemistry technology using the Qwin image analysis system.SIgA postive cells were distributed and located primarily in inhesion membrane and enteraden cavity of the duodenum mucosa,additive IL-2 in the Shigella vaccine could increase secretion of SlgA positive cells and strengthen the immunity of the birds,in some degree.The 50 μg IL-2 in the vaccine could display a optimal immunological enhancement effect.

OBJECTIVETo prepare the cubic phase gel containing capsaicin and characterize its properties.

METHODThe cubic phases gel composed of glycerol monoolein, capsaicin and water was made by self-emulsion technology. The characterization of cubic phase gel was carried out by cross-polarizing light microscopy (CPLM) and Small Angle X-Ray Scattering (SAXS). The capsaicin content was determined by HPLC analysis.

RESULTUnder CPLM, cubic phase gel showed dark background. SAXS scattering spectra showed the scattering peaks at 0.1096, 0.1334, 0.1557, 0.1883 A(-1) which was compatible with q1:q2:q3:q4 = mean square root of 2: mean square root of 3: mean square root of 4: mean square root of 6. It was well known that the scattering vector ratio was the characteristic of cubic phase and the internal structure was confirmed to be Pn3m (Q224). The linear range for capsaicin determination was 3.25 x 10(-4) - 2.08 x 10(-2) g x L(-1) (R2 = 1). The average recovery was 97.53% with RSD of 2.9% (n=9).

CONCLUSIONCPLM and SAXS technology are suitable to characterize the cubic phase gel The determination of the capsaicin content by HPLC is simple and reproducible.

Capsaicin ; chemistry ; Chemistry, Pharmaceutical ; Microscopy, Polarization ; Plant Extracts ; chemistry ; Scattering, Small Angle

OBJECTIVETo investigate the apoptosis inducing effects of ponicidin (PON) on leukemic K562 cells and its mechanisms of action.

METHODK562 cells in culture medium in vitro were given different concentrations of PON (10-50 micromol x L(-1)) for 24, 48 and 72 h. The inhibitory rate of the cells was measured by MTT assay, cell apoptotic rates were detected by flow cytometry (FCM) using Annexin V staining after K562 cells were treated with different concentrations of PON for 72 hours, and cell morphology was observed by Wright-Giemsa staining. Western blot was used to detect caspase-3 and poly(ADP-ribose) polymerase (PARP) expression, and the protein levels in mitogen-activated protein kinase signaling pathways (MAPKs, p-P38, p-ERK and p-JNK) as well as p-AKT and p-P85 in PI3K/AKT signaling pathways were also detected.

RESULTPON (over 30 micromol x L(-1)) could inhibit the growth of K562 cells in both time- and dose-dependent manner. FCM analysis revealed that apoptotic cells were gradually increased in a dose-dependent manner after treatment for 72 hours, and that marked morphological changes of cell apoptosis such as condensation of chromatin was clearly observed by Wright-Giemsa staining after treatment by 50 micromol x L(-1) PON. Western blot showed cleavage of the caspase-3 zymogen protein (32 kD), with the appearance of its 17 kD subunit, and a cleaved 89 kD fragment of 116 kD PARP was also found. Furthermore, Western blotting also showed that expression of p-AKT and p-P85 in PI3K/AKT signaling pathways was downregulated dramatically whereas the expression of p-P38 as well as p-ERK and p-JNK remained unchanged after the cells were treated by PON for 48 h.

CONCLUSIONThe results demonstrate that PON exhibits in vitro anti-leukemia effect by induction of apoptosis in K562 cells, and that PON induced apoptosis in K562 cells mainly related to activation of caspase-3 as well as inactivation of PI3K/AKT signaling pathway via down regulation of the expression of p-AKT and p-P85 protein levels. These results provide strong laboratory evidence for further anti-leukemia trials of PON.

Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Diterpenes ; pharmacology ; Humans ; Poly(ADP-ribose) Polymerases ; metabolism ; Signal Transduction ; drug effects

Objective:To observe the changes in blood flow density of radial retinal peripapillary capillary (RPC) around the optic disc in patients with non-arteritic anterior ischemic optic neuropathy (NAION) at different stages of the continuous course of the disease.Methods:A prospective cohort study. From January to December 2020, 29 cases of 29 eyes of NAION patients diagnosed in the Eye Center of the Second Affiliated Hospital of Zhejiang University School of Medicine were included in the study. Among them, there were 18 males with 18 eyes and 11 females with 11 eyes. The average age was 53.62±6.67 years old. The affected eye underwent routine eye examination and visual field, optic cohenrence tomography angiography (OCTA) examination. Visual field inspection was performed to obtain the average visual mean defect (MD) value. OCTA was used to measure the thickness of the peripapillary retinal nerve flayer (pRNFL) around the optic disc, the whole en face image vessel density (wiVD), intro disc vessel density (diVD), RPC blood flow density around the optic disc, and macular ganglion cell complex (GCC). The course of disease ≤3 weeks was defined as the acute phase; 4-12 weeks was defined as the subacute phase; >12 weeks was defined as the chronic phase. The changes of visual field MD, optic disc RPC blood flow density, pRNFL thickness and macular GCC thickness were observed in the acute, subacute and chronic phases (12-24, >24 weeks). A completely randomized design of variance analysis was used to compare the differences in visual field MD, RPC blood flow density, GCC, and pRNFL thickness in different courses. Pearson correlation analysis was used to analyze the correlation between pRNFL thickness, macular GCC thickness, visual field MD changes and RPC blood flow density around the optic disc sex.Results:The wiVD of the eyes in the acute phase, subacute phase, and chronic phase (12-24 weeks, >24 weeks) were (44.96±2.76)%, (41.50±3.49)%, (39.08±5.43)%, (38.56±6.48)%. There was a statistically significant difference in wiVD of eyes with different disease courses ( F=8.939, P<0.001). The average difference of wiVD between 12-24 weeks and >24 weeks in the chronic phase was -0.984, and the difference was not statistically significant ( P>0.05). There was no statistically significant difference in diVD of patients with different courses of disease ( F=1.079, P=0.365). The blood flow density of RPC around the optic disc of the affected eye, except for the lower part, the blood flow density of the nasal side, the temporal side, and the upper quadrant, decreased significantly with the progression of the disease, and the difference was statistically significant ( F=8.816, 6.069, 8.943; P<0.05). In the chronic phase, the average difference of blood flow density between the nasal, temporal, and upper sides of the eyes between 12-24 weeks and more than 24 weeks in the chronic phase was -0.984, -0.230, -0.198, and the difference was not statistically significant ( P>0.05). There was no statistically significant difference in the visual field MD of patients with different courses of disease ( F=0.277, P=0.842); the overall pRNFL thickness and average macular GCC thickness were compared with statistical significance ( F=47.122, 14.954; P<0.001, <0.001), all became significantly thinner with the progression of the disease. The results of Pearson correlation analysis showed that the blood flow density of the entire optic disc wiVD, the blood flow density of RPC in the temporal quadrant around the optic disc and the visual field MD ( r=-0.225, -0.268; P<0.05), and the average thickness of GCC ( r=0.480, 0.436; P<0.01) were all related. Conclusion:The blood flow density of RPC in the entire optic disc and around the optic disc (except the lower quadrant) of NAION eyes gradually decrease with the progression of the disease, and stabilize after 12 weeks of the disease.

Objective:To observe the clinical features and visual prognostic factors of ethambutol-induced optic neuropathy (EON).Methods:A cohort study. Twenty-four inpatients (46 eyes) identified as EON in Neuro-Ophthalmology Department of Chinese PLA General Hospital from January 2014 to December 2017 were enrolled, including 14 males (26 eyes) and 10 females (20 eyes) with a ratio of 1.4/1 male/female. The average age was 42.79±15.12 years and the average weight was 62.46±12.31 kg. The average time duration between oral administration of ethambutol and occurrence of EON was 9.94±16.49 months. The average time of ethambutol duration was 7.06±11.68 months, with an average accumulative dose of 156.7±1 779.0 g and the average daily dose of 15.07±8.95 mg/(kg·d). All patients were tested with visual acuity, fundus photos, colour vision, OCT, visual field, VEP, orbital MRI and the gene of OPA1 and mitochondrial deoxyribonucleic acid(mtDNA). All the patients accepted drug withdrawal immediately after diagnosis, and were given the treatment of systemic nerve nutrition and improvement of microcirculation for 2 weeks. The time of follow-up was more than 12 months. According to whether the visual acuity (VA) in any of eyes was over than 0.1 at the last follow- up, all the patients were divided into two groups: the bad VA group (VA less than or equal to 0.1) and the better VA group (VA over than 0.1) group. The χ2 test and Fisher's exact probabilistic method test were used to compare the counting data between groups, and the Wlincox rank sum test was used to compare the measurement data. Multiple factors of VA outcome between the patients with bad or better va were analyzed by logistic regression. Results:Thirty eyes (65.2%) had VA less than or equal to 0.1 and 5 eyes (10.9%) had VA over than 0.5 at EON onset. The VA of the rest 11 eyes (23.9%) was higher than 0.1 and lower than 0.5. At the last follow- up, 20 eyes (43.5%) had VA less than or equal to 0.1 and 9 eyes (19.6%) had VA over than 0.5, the VA of the rest 17 eyes (36.9%) was higher than 0.1 and lower than 0.5. Fundus examination revealed 7 eyes (15.3%) with optic disc edema. OCT revealed significant loss of the retinal nerve fiber layer (RNFL) in the affected eyes, mainly in the temporal RNFL of the optic disc. All patients had dyschromasia, mainly in distinguishing the color of red and green. The types of visual field defect was as following: central dark spot (52.2%), diffuse visual acuity decreased (30.4%), temporal hemianopsia (17.4%). Orbital MRI revealed that 12/24 (50.0%) patients had T2 lesions with T1 enhancement in 6/24 patients (25.0%). Genetic test showed that 4 patients (16.7%) had gene mutation. Among them, there were 2 patients with OPA1 mutation, 1 with mtDNA 14340 point mutation and 1 with the mtDNA 11778 point mutation. Thirteen patients showed better VA outcomes (over than 0.1) while 11 showed bad VA outcomes after discontinuation of ethambutol. Between the better VA group and the bad VA group, there were statistically significant differences in the daily dose of ethambutol and gene mutation( P=0.031, 0.023). The daily dose was related to visual prognosis of EON while only the daily dose of more than 18 mg/(kg·d) may lead to bad VA outcomes according to the logistic analysis (95% CI 0.007-0.736, OR=0.069, P=0.027). Conclusions:EON may have OPA1 and mtDNA mutation with more bilateral eyes involved and less optic edema, which about 43.5% of the patients showed irreversible visual impact. The daily dose of ethambutol is related to the vision recovery.

Intranasal drug delivery system is a non-invasive drug delivery route with the advantages of no first-pass effect, rapid effect and brain targeting. It is a feasible alternative to drug delivery via injection, and a potential drug delivery route for the central nervous system. However, the nasal physiological environment is complex, and the nasal delivery system requires "integration of medicine and device". Its delivery efficiency is affected by many factors such as the features and formulations of drug, delivery devices and nasal cavity physiology. Some strategies have been designed to improve the solubility, stability, membrane permeability and nasal retention time of drugs. These include the use of prodrugs, adding enzyme inhibitors and absorption enhancers to preparations, and new drug carriers, which can eventually improve the efficiency of intranasal drug delivery. This article reviews recent publications and describes the above mentioned aspects and design strategies for nasal intranasal drug delivery systems to provide insights for the development of intranasal drug delivery systems.
Administration, Intranasal ; Drug Delivery Systems ; Pharmaceutical Preparations ; Drug Carriers ; Brain ; Nasal Cavity/physiology* ; Nasal Mucosa

The purpose of this study was to compare the pharmacokinetic profiles of tetramethylpyrazine phosphate (TMPP) in plasma and extracellular fluid of the cerebral cortex of rats via three delivery routes: intranasal (i.n.), intragastric (i.g.) and intravenous (i.v.) administration. After i.n., i.g. and i.v. administration of a single-dose at 10 mg/kg, cerebral cortex dialysates and plasma samples drawn from the carotid artery were collected at timed intervals. The concentration of TMPP in the samples was analyzed by HPLC. The area under the concentration-time curve (AUC) and the ratio of the AUCbrain to the AUCplasma (drug targeting efficiency, DTE) was calculated to evaluate the brain targeting efficiency of the drug via these different routes of administration. After i.n. administration, TMPP was rapidly absorbed to reach its peak plasma concentration within 5 min and showed a delayed uptake into cerebral cortex (t max=15 min). The ratio of the AUCbrain dialysates value between i.n. route and i.v. injection was 0.68, which was greater than that obtained after i.g. administration (0.43). The systemic bioavailability obtained with i.n. administration was greater than that obtained by the i.g. route (86.33% vs. 50.39%), whereas the DTE of the nasal route was 78.89%, close to that of oral administration (85.69%). These results indicate that TMPP is rapidly absorbed from the nasal mucosa into the systemic circulation, and then crosses the blood-brain barrier (BBB) to reach the cerebral cortex. Intranasal administration of TMPP could be a promising alternative to intravenous and oral approaches.

Objective:To evaluate the efficacy and safety of oral anisodine hydrobromide tablets in the treatment of nonarteritic anterior ischemic optic neuropathy (NAION).Methods:A multicenter nonrandomized controlled trial was conducted.A total of 282 acute NAION patients (282 eyes) were recruited from 16 hospitals in China from July 2020 to May 2021.Patients were divided into two groups according to treatment methods, which were control group (124 cases, 124 eyes) receiving regular treatment including citicoline sodium plus Ginkgo biloba leaf liquid extract or Ginkgo biloba leaf extract tablets plus mecobalamin, and experimental group (158 cases, 158 eyes) receiving treatment in control group plus oral anisodine hydrobromide tablets 1 mg, twice daily for 2 to 3 months.Best corrected visual acuity (BCVA), visual field index (VFI), peripapillary retinal nerve fiber layer (pRNFL) and radial peripapillary capillary vessel density (RPC) were assessed at 1, 2, 3, and 6 months after enrollment using the standard decimal visual acuity chart, 750i Humphery visual field analyzer, Cirrus HD-OCT 4000/Cirrus HD-OCT 5000, RTVue-XR optical coherence tomography respectively.The primary outcomes were BCVA and VFI, and the secondary outcomes were pRNFL, RPC, and the side effects during the follow-up.The study adhered to the Declaration of Helsinki.All patients were fully informed about the treatment and purpose of this study and voluntarily signed the informed consent form.The study protocol was approved by Chinese PLA General Hospital (No.S2020-021-01). Results:In all, 242 patients (242 eyes) completed the follow-up of BCVA, and 98 patients (98 eyes) completed the VFI follow-up.In terms of visual function, BCVA and VFI improved significantly over time in the two groups, and BCVA and VFI were better in experimental group than in control group at various follow-up time points (all at P<0.05). In terms of structure, pRNFL gradually decreased in both groups with the extension of treatment, and pRNFL was significanthy thinner in experimental group than in control group at various follow-up time points (all at P<0.05). There was no significant difference in RPC between the two groups at the last follow-up ( P>0.05). There were two cases with side effects and one case was discontinued due to side effects 25 days after enrollment. Conclusions:Oral anisodine hydrobromide can improve visual acuity and visual field in NAION and accelerate the regression of optic disc edema, with good safety.

An explicit illustration of pulmonary delivery processes (PDPs) was a prerequisite for the formulation design and optimization of carrier-based DPIs. However, the current evaluation approaches for DPIs could not provide precise investigation of each PDP separately, or the approaches merely used a simplified and idealized model. In the present study, a novel modular modified Sympatec HELOS (MMSH) was developed to fully investigate the mechanism of each PDP separately in real-time. An inhaler device, artificial throat and pre-separator were separately integrated with a Sympatec HELOS. The dispersion and fluidization, transportation, detachment and deposition processes of pulmonary delivery for model DPIs were explored under different flow rates. Moreover, time-sliced measurements were used to monitor the PDPs in real-time. The Next Generation Impactor (NGI) was applied to determine the aerosolization performance of the model DPIs. The release profiles of the drug particles, drug aggregations and carriers were obtained by MMSH in real-time. Each PDP of the DPIs was analyzed in detail. Moreover, a positive correlation was established between the total release amount of drug particles and the fine particle fraction (FPF) values ( = 0.9898). The innovative MMSH was successfully developed and was capable of illustrating the PDPs and the mechanism of carrier-based DPIs, providing a theoretical basis for the design and optimization of carrier-based DPIs.