Ability of Tong Luo Jiu Nao Oral Solution to Improve Cognitive and Gait Deficits in a Rat Model of Focal Cerebral Ischemia

Objective To investigate the ability of Tong Luo Jiu Nao(TLJN)Oral Solution to improve cognitive and gait deficits in a rat model of focal cerebral ischemia.Methods An intraluminal suture method was used to create a rat model of focal cerebral ischemia.Experimental animals were divided into4groups:sham surgery,cerebral ischemia model,cerebral ischemia model+low-dose TLJN Oral Solution(150mg/kg),cerebral ischemia model+high-dose TLJN Oral Solution(300mg/kg).After4weeks of drug intervention,water maze and gait analyses were conducted.Brain tissue was stained with hematoxylin and eosin and used to monitor and calculate the volume ratio of cerebral infarction in each group of permanent middle cerebral artery occlusion(pMCAO)rats.Results(1)Gait analyses:TLJN Oral Solution significantly improved the overall gait of the focal cerebral ischemic rat model.The improvement in walking speed might have been achieved by the shortening of the animals’stride time.The shortening of stance time by TLJN in the cerebral ischemic animals might be due to the shortening of brake and propulsion times,while the shortening of stride time by TLJN in the cerebral ischemic animals might have been due to the shortening of stance time and swing time.(2)Water maze navigation test:high-dose TLJN Oral Solution significantly reduced the animals’total swimming distance and duration and increased the proportion of swimming distance and time spent in the target quadrant,suggesting that TLJN significantly improved the cognitive deficit in the animal model.(3)High-dose TLJN significantly reduced the cerebral infarction volume ratio of the pMCAO rats.Conclusion This intraluminal suture method can reliably replicate a rat model of focal cerebral ischemia.TLJN Oral Solution improves the gait and cognitive function of the focal cerebral ischemia rat model to various extents.

Effects of dielectric decrement on surface potential in a mixed electrolyte solution

Surface potential is an important parameter related to the physical and chemical properties of charged particles. A simple analytical model for the estimation of surface potential is established based on the Poisson–Boltzmann theory with the consideration of the dielectric decrement in mixed electrolyte. The analytical relationships between surface potential and charge density are derived in different mixed electrolytes with monovalent and bivalent ions. The dielectric decrease on the charged surface strongly affects the surface potential at a high charge density with different ion strengths and concentration ratios of counter-ions. The surface potential based on the Gouy–Chapman model is underestimated because of the dielectric decrement on the surface. The diffuse layer can be regarded as a continuous uniform medium only when the surface charge density is lower than 0.3 C·m-2. However, the surface charge densities of many materials in practical applications are higher than 0.3 C·m-2. The new model for the estimation of surface potential can return to the results obtained based on the Gouy–Chapman model at a low charge density. Therefore, it is implied that the established model that considers the dielectric decrement is valid and widely applicable.

Principles of Surface Potential Estimation in Mixed Electrolyte Solutions:Taking into Account Dielectric Saturation

The dielectric properties between in-particle/water interface and bulk solution are significantly different,which are ignored in the theories of surface potential estimation.The analytical expressions of surface potential considering the dielectric saturation were derived in mixed electrolytes based on the nonlinear Poisson-Boltzmann equation.The surface potentials calculated from the approximate analytical and exact numerical solutions agreed with each other for a wide range of surface charge densities and ion concentrations.The effects of dielectric saturation became important for surface charge densities larger than 0.30 C/m^2.The analytical models of surface potential in different mixed electrolytes were valid based on original Poisson-Boltzmann equation for surface charge densities smaller than 0.30 C/m^2.The analytical model of surface potential considering the dielectric saturation for low surface charge density can return to the result of classical Poisson-Boltzmann theory.The obtained surface potential in this study can correctly predict the adsorption selectivity between monovalent and bivalent counterions.

Effect of different modeling time on intestinal bacteria in letrozole induced PCOS rats

Objective:The differences of ovarian morphology,reproductive hormones,glucose and lipid metabolism and intestinal bacteria in rats with polycystic ovary syndrome(PCOS)induced by triazole were compared.Method:Eighteen 21 SPF female SD rats were randomly divided into group A(3-week group),group B(5-week group)and group D(control group)by random number table.Group A received letrozole+CMC-Na mixture by gavage in the first 3 weeks and CMC-Na solution by gavage in the last 2 weeks,group B received letrozole+CMC-Na mixture by gavage for 5 weeks,and group D received CMC-Na solution by gavage for 5 weeks,and all three groups of rats were fed with normal diet.At the end of gavage,the body weight of rats in each group was observed,the histological changes of ovaries were observed by hematoxylin-eosin(HE)staining,the serum levels of estradiol(E2),follicle stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),total cholesterol(TC),triglyceride(TG),fasting blood glucose(Glu),fasting insulin(FINS)and lipopolysaccharide(LPS)were measured by enzyme-linked immunosorbent assay(ELISA),and the LH/FSH ratio and insulin resistance index(HOMA IR)were calculated;the intestinal bacteria of rats were detected by 16S rRNA technique.Result:1.Comparison of ovary histomorphology:Under light microscope,multiple luteum and oocytes were observed in mature follicles in group D,and granulosa cells were orderly arranged and multilayered,without cystic dilated follicles.There were no mature follicles in the ovarian tissues of group A and GROUP B.The follicles were irregular in structure and more cystic dilated follicles were visible.The number of granular cells in some follicles decreased or even disappeared.2.Comparison of sex hormone levels:compared with group D,T level in group B was significantly increased(P<0.001),and T level in group A had an upward trend(P>0.05);The LH/FSH levels in group A and B were significantly increased(P<0.001;P<0.001).Compared with group A,E2 in group B was significantly decreased(P<0.05)and T was significantly increased(P<0.01).3.Comparison of glucose and lipid metabolism levels:Compared with group D,TC levels in groups A and B were significantly increased(P<0.01;P<0.01).Compared with group A,TG in group B was significantly increased(P<0.05).There were no significant differences in Glu,FINS and HOMA-IR levels among all groups.4.Comparison of LPS levels:Compared with group D,the serum LPS levels of rats in groups A and B were significantly increased(P<0.001;P<0.01).5.Intestinal flora analysis and comparison:At the phylum level,compared with group D,the abundance of Firmicutes in group B increased(P<0.01),Firmicutes in group A showed an upward trend(P>0.05),and the abundance of Bacteroidetes in groups A and B decreased(P<0.05).At the genus level,compared with group D,Lactobacillus in group B increased(P<0.01).The results of LEfSe analysis showed that there were differences in the composition of various intestinal bacteria among the three groups(LDA>3).Conclusion:The phenotype of PCOS rats was related to the length of modeling,and the phenotypic characteristics of PCOS in rats at 5 weeks of modeling were more typical than those in rats at 3 weeks of modeling;PCOS can cause changes in intestinal flora,and the changes in the structure of intestinal flora between groups are related to different modeling duration.

Antidepressant-like effects and memory enhancement of a herbal formula in mice exposed to chronic mild stress

Shen Yuan Gan（SYG） is a Chinese herbal prescription composed of total saponins of Panax ginseng and total oligosaccharide esters of Polygala tenuifolia（2：1）.Our previous studies have demonstrated that SYG has antidepressant-like effects in various mouse models of behavioral depression.The present study aimed to test whether SYG affected chronic mild stress（CMS）-induced depression and cognitive impairment in mice.We found that a 5-week CMS schedule induced significant degradation of the coat state,decreased sucrose intake in the sucrose-preference test,and increased the latency to feed in the noveltysuppressed feeding test.All of these CMS-induced changes were ameliorated by SYG（100 and 200 mg/kg） and fluoxetine（10 mg/kg）.In addition,SYG restored the decreased monoamine neurotransmitter concentrations（serotonin,dopamine,norepinephrine and acetylcholine） induced by CMS in the prefrontal cortex.Interestingly,SYG ameliorated CMS-induced cognitive impairment in the step-through test,and increased the acetylcholine level in the prefrontal cortex.These results suggest that SYG has an antidepressant-like action and enhances cognition by modulating the serotonin,dopamine,norepinephrine,and acetylcholine levels in the prefrontal cortex.

Acupuncture versus oral medicine for women with diminished ovarian reserve:A cohort study

Objective:To observe whether electro-acupuncture(EA) is better in improving hormone levels among women with diminished ovarian reserve(DOR) compared with oral medicine [Chinese herbs with or without hormone replacement therapy(HRT)] that is often used in gynecology clinics in China.Methods:Eligible participants entered the EA group or medication group according to their preferences.Participants in the EA group received EA treatment,and participants the medication group received herbal decoction alone or combined with HRT.Reproductive hormone levels of the participants were tested before and during treatment in both groups.Results:A total of 80 women with DOR were screened and 57 cases were included(29 in the EA group and 28 in the medication group) in the final analysis.The differences in the change from baseline between the EA and medication group in mean follicle-stimulating hormone(FSH) levels were-6.6(95% Cl,-17.03 to 3.72),(P=0.848) and-7.09(95% CI,-15.86 to 1.66),(P=0.539) at weeks 12 and 24,respectively.The proportion of women with regular FSH levels in the EA and medication group was 51.3% vs 32.1% at week 12(P=0.134) and 51.3% vs 25.0% at week 24(P=0.038).Conclusion:EA might influence regulating the levels of FSH,luteinizing hormone(LH),estrogen and the FSH/LH ratio,and no difference was found between the effects of EA and oral medicine in changes of reproductive hormone levels.

Cognitive-enhancing effects of hydrolysate of polygalasaponin in SAMP8 mice

Objectives： The aim of the study is to evaluate the cognitive-enhancing effects of hydrolysate of polyga- lasaponin （HPS） on senescence accelerate mouse P8 （SAMP8） mice, an effective Alzheimer＇s disease （AD） model, and to research the relevant mechanisms. Methods： The cognitive-enhancing effects of HPS on SAMP8 mice were assessed using Morris water maze （MWM） and step-through passive avoidance tests. Then N-methyl-D-aspartate （NMDA） receptor subunit expression for both the cortex and hippocampus of mice was observed using Western blot- ting. Results： HPS （25 and 50 mg/kg） improved the escape rate and decreased the escape latency and time spent in the target quadrant for the SAMP8 mice in the MWM after oral administration of HPS for 10 d. Moreover, it decreased error times in the passive avoidance tests. Western blotting showed that HPS was able to reverse the levels of NMDAR1 and NMDAR2B expression in the cortex or hippocampus of model mice. Conclusions： The present study suggested that HPS can improve cognitive deficits in SAMP8 mice, and this mechanism might be associated with NMDA receptor （NMDAR）-related pathways.

Effects of Zhengtian Pills on Migraine Headache in Rats via Transient Receptor Potential Vanilloid 1

Objective To investigate the effects and molecular mechanism of Zhengtian Pills （ZTP） on migraine headache. Methods All rats were randomly divided into control, positive control, migraine model, low- and high-dose ZTP groups, and glyceryl trinitrate was injected to induce migraine headache. The time of ears turning red, frequency of scratching head, climbing the cage, and head-twitching were used to evaluate rat behaviors. After 10 d administration of ZTP, the expression levels of transient receptor potential vanilloid 1 （TRPV1） both in cortex and hippocampus were determined by Western blotting. Results After 2 min of glyceryl trinitrate injection, rats showed headache phenomena that parallels the clinical symptoms of migraine, which peaked in 30 min, and lasted for 60 min. Frequency of head-twitching and numbers of scratching head in glyceryl trinitrate （GTN） group were significantly increased. In contrast, after ZTP （1.08 g/kg, ig） treatment, the numbers of scratching head with fore-limb, hind-limb and the frequency of head-twitching were significantly decreased. Flunarizine （FLU） and low-dose ZTP （0.54 g/kg） also showed a trend to decrease the numbers of scratching head and head-twitching frequency, but no significant difference. Besides, ZTP significantly decreased the up-regulated TRPV1 protein expression level both in cortex and hippocampus. Conclusion The present study shows that ZTP could significantly improve the migraine symptoms of headache in rats and TRPV1 might be one of the important molecular mechanisms. This is the first report about the effect of ZTP on TRPVl protein expression level both in cortex and hippocampus of rats.

Significance of alternative splicing in cancer cells

Objective:Alternative splicing can generate various structural and functional protein isoforms.Recently,accumulating evidence shows a relationship between alternative splicing and cancer.Cancer is a complex and chronic disease that involves malignant transformation.In this review,we consider alternative splicing events in relation to the hallmarks of cancer cells,and discuss current therapies to treat cancer-related to alternative splicing.Data sources:Data cited in this article are from the PubMed and Embase database,primarily focusing on research published from 2000 to 2018.Study selection:Articles were selected with the search terms“alternative splicing,”“cancer cell,”“tumor microenvironment,”and“therapy.”Results:Alternative splicing plays an important role in tumorigenesis,development,and escape from cell death.Taking this trait of cancer cells into consideration will allow more definite diagnoses of cancer,and allow the development of more effective medicines to intervene in cancer that could focus on controlling alternative splicing or competitively binding to the final products.Conclusions:Alternative splicing is common in cancer cells.Consideration of alternative splicing may allow different strategies for cancer therapy or the identification of novel biomarkers for cancer diagnosis.

Paraneoplastic Eosinophilia in Clear Cell Renal Cell Carcinoma

To the Editor：Eosinophilia commonly occurs secondary to parasitic infections,allergy,collagen-vascular disease,hypersensitivity conditions,or underlying hematologic or solid malignancies.However,eosinophilia associated with clear cell renal cell carcinoma （CCRCC） is rare.Herein,we report a case of paraneoplastic eosinophilia associated with CCRCC.

Neuroprotection of Stilbenes from Leaves of Cajanus cajan against Oxidative Damage Induced by Corticosterone and Glutamate in Differentiated PC12 Cells

Objective To investigate the neuroprotective effects of four stilbenes, namely cajaninstilbene acid （CSA）, Iongistyline A, Iongistyline C, and cajanolactone A, isolated from the leaves of Cajanus cajan. Methods Neuroprotective effects of the four stilbenes were evaluated using rat pheochromocytoma cell line （PC12 cells） damage models induced by corticosterone （Cort） or glutamate. In order to elaborate whether the neuroprotective effects of stilbenes are related to anti-oxidant properties, both oxidant and anti-oxidant parameters were measured. Results The results of MTT assay and LDH release assay demonstrated that the four stilbenes possessed neuroprotective effects. Moreover, the treatment on PC12 cells with Cort or glutamate （Glu） could significantly increase the levels of ROS and MDA with decreasing the activities of SOD and CAT. However, the four tested stilbenes could significantly alleviate such situation by dropping out the levels of ROS and MDA, as well as enhancing the activities of SOD and CAT. Conclusion These results provide a scientific basis for further studies to explore the potential neuroprotective effects on neurodegenerative diseases.

Zhengtian Pills accelerated long term potentiation both in Schaffercollateral-CA1 and perforant path-dentate gyrus synapses

Objective: To investigate the effects of Zhengtian Pills(ZTP) on long term potentiation(LTP) both in Schaffer-CA1 in vitro and perforant path-dentate gyrus(PP-DG) synapses in vivo.Methods: Sprague-Dawley rats were randomly divided into five groups: control, positive control, migraine model, low-, and high-dose ZTP groups. Glyceryl trinitrate(10 mg/kg) was injected subcutaneously to make migraine rat model. Flunarizine(0.9 mg/kg) was set as positive control. Extracellular recording technique in vivo was used to record the effects of ZTP on LTP of PP-DG pathway in anesthetized rats; Using extracellular recording technique in vitro, the effects of ZTP on LTP of Schaffer Collateral-CA1 pathway in rat hippocampal slices were investigated.Results: Compared to the control group, ZTP(1.08 g/kg) significantly enhanced population spike amplitude in PP-DG pathway; Glyceryl trinitrate(10 mg/kg) significantly reduced population spike amplitude in PP-DG pathway; Neither ZTP(0.54 g/kg) nor Flunarizine(0.9 mg/kg) had significant effects on LTP in PP-DG pathway. Compared to the model group, ZTP(1.08 g/kg), ZTP(0.54 g/kg), and flunarizine(0.9 mg/kg) significantly enhanced population spike amplitude in PP-DG pathway. Compared to the control group, ZTP(1.08 g/kg) significantly enhanced field excitatory postsynaptic potential(fEPSP) slope in Schaffer Collateral-CA1 pathway; Glyceryl trinitrate(10 mg/kg) significantly reduced f EPSP slope in Schaffer Collateral-CA1 pathway; Neither ZTP(0.54 g/kg) nor flunarizine(0.9 mg/kg) significant effects on LTP in Schaffer Collateral-CA1 pathway, Compared to the model group, ZTP(1.08 g/kg), ZTP(0.54 g/kg), and Flunarizine(0.9 mg/kg) had significantly enhanced fEPSP slope in Schaffer Collateral-CA1 pathway.Conclusion: Combined with the previous study, the results gave a clue that the effects of ZTP on TRPV1 and hippocampal LTP or their interactions could be the important molecular mechanisms of ZTP acting as migraine and headache medication.

Screening of Eleven Commonly Used Traditional Chinese Medicines for Inhibitory Effects on Human Cytochrome P450 Enzymes

Objective Traditional Chinese medicines（TCMs） as natural therapeutic agents have been widely used and received great attention over the world.However,the information on them to cause herb-drug interactions mediated by cytochrome P450（CYP） enzymes is limited.The present study aims to evaluate the inhibitory effects of 11 commonly used TCMs,including Nelumbinis Folium,Ginseng Radix et Rhizoma,Ziziphi Spinosae Semen,Chrysanthemi Flos,Lonicera Japonica Flos,Lycii Fructus,Cassiae Semen,Poria,Crataegi Fructus,Schisandrae Chinensis Fructus,and Polygonati Rhizoma,on the five human major CYP isoenzymes 2C19,2D6,3A4,2E1,and 2C9.Methods An in vitro cocktail method was used.Results The aqueous extracts of all tested TCMs were found no significant inhibitory effect,with IC50 values higher than 100 μg/mL while,the ethanolic extracts of some herbs showed more potent inhibition.These herbs include Nelumbinis Folium with IC50 values of 1 2.05 and 61.43 μg/mL on CYP2D6 and CYP2E1,Schisandrae Chinensis Fructus with IC50 values of 64.42 and 47.24 μg/mL on CYP2C1 9and CYP3A4,and Chrysanthemi Flos with IC50 values of 45.25 μg/mL on CYP2C9,respectively.Conclusion Co-administration of these TCMs and their products with conventional medicines should be paid much attention to,and their potential inhibitory effects on CYP enzyme activities need to be further investigated.