Structural diversification of bioactive bibenzyls through modular co-culture leading to the discovery of a novel neuroprotective agent

Bibenzyls, a kind of important plant polyphenols, have attracted growing attention for their broad and remarkable pharmacological activities. However, due to the low abundance in nature, uncontrollable and environmentally unfriendly chemical synthesis processes, these compounds are not readily accessible. Herein, one high-yield bibenzyl backbone-producing Escherichia coli strain was constructed by using a highly active and substrate-promiscuous bibenzyl synthase identified from Dendrobium officinale in combination with starter and extender biosynthetic enzymes. Three types of efficiently post-modifying modular strains were engineered by employing methyltransferases, prenyltransferase, and glycosyltransferase with high activity and substrate tolerance together with their corresponding donor biosynthetic modules. Structurally different bibenzyl derivatives were tandemly and/or divergently synthesized by co-culture engineering in various combination modes. Especially, a prenylated bibenzyl derivative ( 12) was found to be an antioxidant that exhibited potent neuroprotective activity in the cellular and rat models of ischemia stroke. RNA-seq, quantitative RT-PCR, and Western-blot analysis demonstrated that 12 could up-regulate the expression level of an apoptosis-inducing factor, mitochondria associated 3 (Aifm3), suggesting that Aifm3 might be a new target in ischemic stroke therapy. This study provides a flexible plug-and-play strategy for the easy-to-implement synthesis of structurally diverse bibenzyls through a modular co-culture engineering pipeline for drug discovery.

Relationship between fragmented QRS wave with ventricular arrhythmia's incidence situation and heart rate variability in patients with old myocardial infarction / 中国综合临床

Objective:To explore the relationship between fragmented QRS complex and heart rate variability (HRV) and ventricular arrhythmia in patients with old myocardial infarction.Methods:From August 2018 to October 2019, 200 patients with old myocardial infarction were first treated in the Department of cardiac function examination of the First Affiliated Hospital of Hebei North University. The patients were divided into 99 cases of old myocardial infarction with fragmented QRS wave group and 101 cases of old myocardial infarction without fragmented QRS wave group according to the case bank data and conventional 12 lead ECG diagnosis in our hospital for the first time. Then, the 24-h ambulatory ECG reexamined within 1 year after discharge was retrospectively analyzed. The incidence of ventricular arrhythmia was compared between the two groups by χ 2 test. The difference of heart rate variability between the two groups was compared by rank sum test. Multiple logistic regression was used to analyze the value of different indexes of heart rate variability in the evaluation of fragmented QRS complex in old myocardial infarction. Drawing the receiver operating characteristic (ROC), and the area under the curve (AUC) was used to analyze the diagnostic accuracy of different indexes of heart rate variability in the broken QRS complex of old myocardial infarction. Results:According to the Lown classification of ventricular premature contraction, the number of positive ventricular arrhythmias in patients with Grade Ⅰ of ventricular premature contraction and Grade Ⅲ-Ⅴ of ventricular premature contraction in the old myocardial infarction fragmented QRS group was higher than that in the old myocardial infarction non fragmented QRS group (Grade Ⅰ of ventricular premature contraction: 54.5% (54/99)and 39.6%(40/101); χ 2=4.484, P<0.05;Grade Ⅲ-Ⅴ of ventricular premature contraction: 34.3% (34/99) and 9.9%(10/101); χ 2=17.406, P<0.05)). Ventricular premature contraction Grade 0 old myocardial infarction fragmented QRS group was lower than old myocardial infarction non fragmented QRS group (8.1% (8/99) and 48.5% (49/101); χ 2=37.995, P<0.05). The total number of positive cases of ventricular arrhythmia in the old myocardial infarction group with fragmented QRS wave was higher than that in the old myocardial infarction group without fragmented QRS wave (91.9% (91/99) and 51.5%(52/101); χ 2=57.146, P<0.05)). There was no significant difference in the number of positive ventricular arrhythmias between the old myocardial infarction fragmentation QRS group and the old myocardial infarction non fragmentation QRS group ( P>0.05). The standard deviation of NN intervals (SDNN) and the standard deviation of average NN intervals (SDANN) of HRV time domain indexes in the old myocardial infarction fragmented QRS group were higher than those in the old myocardial infarction non fragmented QRS Group (SDNN:143.00(122.00,166.00) vs. 110.00(95.00,130.50), Z=5.780, P<0.05; SDANN:112.00(100.00,136.00) vs. 96.00(76.00,118.50), Z=4.013, P<0.05). Multiple Logistics regression analysis results of HRV domain shows that HRV time domain SDNN and SDANN have diagnositic value in diagnosis fQRS after OMI(SDNN: OR=0.949, 95% CI:0.922-0.977, P<0.001; SDANN: OR=1.036, 95% CI:1.005-1.068, P=0.022). Area under ROC curve of HRV time domain SDNN and SDANN have particular diagnositic accuracy in diagnosis fQRS after OMI(SDNN: AUC 0.737, 95% CI 0.666-0.807, Sensitivity 0.818, Specificity 0.634; SDANN: AUC 0.664, 95% CI 0.587-0.741, Sensitivity 0.737, Specificity 0.673. 0.5<AUC<1). Conclusion:Fragmented QRS complex was positively correlated with the incidence and severity of ventricular arrhythmia in patients with old myocardial infarction, and positively correlated with time-domain indexes SDNN and SDANN of heart rate variability in patients with old myocardial infarction.

TACE using lobaplatin and arsenic trioxide for the treatment of primary hepatic carcinoma in elderly patients: observation of its clinical efficacy / 介入放射学杂志

Objective To investigate the clinical efficacy and safety of transcatheter arterial chemoembolization (TACE) by combination use of lobaplatin and arsenic trioxide in treating primary hepatocellular carcinoma (PHC) in elderly patients.Methods Based on the different medication program,a total of 95 PHC patients,whose liver function belonged to Child-Pugh A or B grade (middle-late stage PHC) or who suffered from early-stage PHC and were unwilling to undergo surgical treatment,were divided into the observation group (n=48) and the control group (n=47).TACE was performed in all patients of both groups.Lobaplatin (40 mg/m2) and arsenic trioxide (10 mg/m2) were adopted for patients of the observation group,while arsenic trioxide (10 mg/m2) was employed for patients of the control group.TACE was carried out once every 6 weeks.The objective response rate (ORR),disease control rate (DCR),the median progression free survival time (mPFS) and the incidence of adverse reactions of both groups were analyzed.Results The ORR of the observation group and the control group was 50.0％ and 48.9％ respectively,and the difference was not statistically significant (P＞0.05).The DCR of the observation group and the control group was 85.4％ and 80.9％ respectively,and the difference was not statistically significant (P＞0.05).The mPFS of the study group and the control group was 9 months and 6 months respectively,and the difference was statistically significant (P＜0.001).The main adverse reactions in the two groups were nausea,vomiting,fever,elevation of aminotransferase,etc.,but the differences between the two groups were not statistically significant (P＞ 0.05).Conclusion For the treatment of PHC in elderly patients,TACE by combination use of lobaplatin and arsenic trioxide can prolong mPFS,and the adverse reactions can be well tolerated by patients.This therapeutic regimen may be a better treatment means for PHC in elderly patients.However,large sample randomized and controlled studies are needed to further confirm its curative effect before it can be reliably used as a routine clinical medication regimen.

FoxOs and osteoporosis / 中国药理学通报

As one subclass of forkhead proteins, the forkhead box O ( FoxO) transcription factors take part in a series of bio-logical processes including cellular apoptosis, damaged DNA re-pair and cleavage of reactive oxygen species(ROS). Increasing evidence highlights that oxidative stress elicited by FoxOs con-tributes to imbalance of redox status in cells related to bone me-tabolism, resulting in development of the pathogenesis of osteo-porosis. This article reviews the relationship of FoxOs and osteo-porosis, which may be beneficial for the research of pathological mechanism and therapeutic strategy of osteoporosis.

Preventive effect of Polygonum Multiflorum on deteriorated micro-structure and biomechanical properties induced by prednisone / 中国药理学通报

Aim To investigate the preventive effect of Polygonum Multiflorum (PM)on the deteriorated mi-cro-structure and biomechanical properties induced by prednisone.Methods Ninety 6-month-old male Sprague-Dawley rats were randomly divided into nine groups,which were control,prednisone,CAL,30%ethanol eluent of the PM(H,M,L),PM(H,M,L). Prednisone was gavaged to rat for 21 weeks as model group of osteoporosis.Meanwhile,tested herbal ab-stract were orally administrated to the modeled rats in-duced by prednisone.At the end of the experiment, the right femur was collected for micro-CT scanning, three-dimensional reconstruction and biomechanical test.Results Compared with the control group,mod-el group showed destruction of bone microarchitecture, BV /TV fell 28.6%(P <0.05),bone biomechanical parameters decreases,and stiffness fell 29.7%(P <0.01 ). Compared with the model group, positive group had significantly improved effect on bone micro-architecture,and biomechanical parameters,BV /TV increased 46.7%(P <0.01 ),and stiffness increased 25.9%(P <0.01 ).30% ethanol eluent of the PM (M,L)dose may improve bone microstructure by in-creasing BV /TV 46.7% (P <0.01 ),40.0% (P <0.05)respectively,PM(H)may improve the biome-chanical parameters by increasing stiffness 24.7%(P<0.05),and 30% ethanol eluent of the PM(H)and PMhigh-dose may improve the biomechanical parame-ters,but not as positive group.Conclusions Predni-sone reduces biomechanical properties of rat femur and deteriorates femoral microstructure.30% ethanol eluent of the PM(M,L)and PM(H)plays a preventive role in the changes of micro-structural and biomechanical properties by prednisone,and increases BMD,whereas other groups have no significant preventive effect.

The expression of long non-coding RNA AK089560 in mesenchymal stem cells undergoing osteogenic and adipogenic differentiation / 中国组织工程研究

BACKGROUND:Recent studies have found that stem cellpluripotency and differentiation is regulated by many long non-coding RNAs (LncRNAs). The expression and effect of LncRNA AK089560 during differentiation of stem cells is unclear. OBJECTIVE:To investigate the expression of LncRNA AK089560 in mesenchymal stem cells C3H10T1/2 undergoing osteogenic and adipogenic differentiation. METHODS:Osteogenic differentiation of mesenchymal stem cells C3H10T1/2 was induced by recombinant human bone morphogenetic protein-2 and evaluated using alkaline phosphatase staining. The adipogenic differentiation of mesenchymal stem cells C3H10T1/2 was induced by three factors (dexamethasone, indomethacin and insulin) and evaluated by oil red O staining. The dynamical expression of LncRNA AK089560 was detected by qRT-PCR assay. The AK089560 secondary structure was predicted using RNAfold software. The relationship between AK089560 and neighboring protein-coding genes was analyzed using UCSC genome browser and visualized by fancyGENE online software. RESULTS AND CONCLUSION:Over 70%of C3H10T1/2 cells were positive for alkaline phosphatase after osteogenic induction and more than 80%of the cells positive for oil red O staining after adipogenic induction. qRT-PCR results showed that the expression of LncRNA AK089560 at days 2, 4, 6 of both osteogenic and adipogenic differentiation was significantly decreased compared with the control group (P<0.05). Bioinformatics analysis showed that there was a stem-loop structure for AK089560 and sense overlap relationship between AK089560 and protein-encoding gene Sema3a. These findings indicate that LncRNA AK089560 expression is reduced during osteogenic differentiation and adipogenic differentiation, showing that AK089560 may be involved in regulating the multi-directional differentiation of stem cells.

Inhibitory effect of berberine on differentiation of rat bone marrow mesenchymal stem cells to adipocytes / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of berberine on differentiation of rat bone marrow mesenchymal stem cells (MSCs) to adipocytes and its mechanism. METHODS Rat MSCs were isolated and cultured, adipocytic differentiation was induced with adipogenesis-inducing medium (AIM). Cells were assigned into 6 groups:normal control, AIM group, AIM+berberine 0.1, 0.3, 1 and 3 μmol·L-1 groups, respectively. Morphology characteristics of mesenchymal stem cells were observed under an inverted microscope and adipocyte levels were analyzed by oil O staining. Alkaline phosphatase (ALP) activity was detected using p-nitrophenyl phosphate as a substrate. The cell survival was determined by MTT assay. Expressions of peroxisome proliferator activated receptor γ (PPARγ), fatty acid binding protein (aP2) and CCAAT enhancer-binding protein α (C/EBPα) mRNA were detected by semiquantitative RT-PCR. RESULTS Compared with normal control group, MSCs adipogenic differentiation, PPARγ, aP2 and C/EBPα mRNA expression significantly increased in AIM group (P<0.01), ALP activity in AIM group significantly decreased (P<0.01). Compared with AIM group, berberine inhibited MSCs adipogenic differentiation (P<0.01) and berberine 0.1, 0.3, 1 and 3 μmol·L-1 increased ALP activity by 26%, 54%, 81% and 122%, respectively. Berberine 3 μmol·L-1 significantly downregulated PPARγ expression (0.91±0.10 vs 1.34±0.06) (P<0.01), aP2 (1.05±0.10 vs 1.53±0.09) (P<0.01) and C/EBPα mRNA (1.24±0.06 vs 1.54±0.09) (P<0.01). Berberine had no effect on proliferation of MSCs. CONCLUSION Berberine inhibits differentiation of MSCs into adipocytes, which might be closely related to the downregulation of PPARγ, aP2 and C/EBPα mRNA.

Regulating effects of Wnt signaling pathway on differentiation of bone marrow mesenchymal stem cells into osteoblasts and adipocytes / 中国组织工程研究

BACKGROUND:Disequilibrium of proportion of adipogenesis and osteogenesis from bone marrow mesenchymal stem cells (BMSCs)is associated with many bone diseases.However,it has been demonstrated that Wnt signaling pathway could play an important role in regulation of BMSC differentiation.OBJECTIVE:To investigate the different gene expression profiles and to find the target gene on Wnt signaling pathway of the BMSCs,after being induced to osteoblasts and adipocytes respectively using Wnt signaling pathway PCR array.METHODS:The third-passage BMSCs,after being induced to osteoblasts and adipocytes respectively for 7 days.The total mRNA of MSCs was extracted by Trizol.BMSC morphology was observed following osteogenic and adipogenic induction under an inverted microscope.Gene array was detected by rat Wnt signaling pathway PCR array.Non-induction group served as controls.The ratio of increase/reduction gene of osteoblasts and adipocytes was calculated.RESULTS AND CONCLUSION:Under an inverted microscope,BMSCs with high homogenicity were obtained following passage 3.BMSCs differentiated into osteoblasts following osteogenic induction,and into adipocytes following adipogenic induction.Compared with non-induction group,fifteen genes(Dkk1,kremen,FZD1,FZD7,et al.)were expressed up-regulated(ratio ＞ 2)and 16(sFrp 5,β-catenin,Dvl3,Tcf7,et al.)genes down-regulated(ratio ＜ 0.5)when the third-passage BMSCs were induced to adipocytes.Six genes(Dkk1,kremen,β-catenin,Wnt11,et al.)were expressed up-regulated and 15 genes(sFrp5,sFRP4,Fzd1,et al.)down-regulated when BMSCs being induced to osteoblasts.Above-mentioned results suggested that Wnt signaling pathway plays an important role in the osteoblast and adipocyte differentiation from BMSCs.

Manufacture and experimental study of the categorizing pedestal of multi-function apparatus in the supply room / 中国实用护理杂志

Objective To control the nosocomial infection and avoid the second pollution of thereclaimed medical apparatus to the washing area of supply room. Methods Count, classification and place of the reclaimed medical apparatus went along at the same self-designed categorizing pedestal of the muhi-function apparatus.No absorbing cloth was needed on the table-board, the dirty water flew into the outfall of the sewer directly and automatically, the discreteness could be unpicked, washed and lis-terized after work. Results It avoided the collision of the apparatuses, shortened the time of counting effectively and reduced the chance of pollution.It also avoided the waste of the manpower, the material resources, the time and the second environmental pollution. The effects of the two table-boards after disinfection had statistical difference (P＜0.01). Conclusions The categorizing pedestal of the mul-ti-function apparatus makes the reclaim and handling of the polluted apparatus more standard and to re-move the pollution more thoroughly.

Protective effects of a combination of estrogen and progestin on ovariectomized rats with osteoporosis / 中国组织工程研究

BACKGROUND: It has been reported that a combination of estrogen and progestin has a protective synergistic effect on osteoporosis with only little side effects.OBJECTIVE: This study was designed to investigate the effect of a combination of norethisterone and ethinyl estradiol (EE) on bone mass in ovariectomized rats.DESIGN: This study was a randomized controlled experiment.SETTING:It was conducted at the Department of Pharmacology of Guangdong Medical University.MATERIALS: Twenty-four specific pathogen free (SPF) unmated SD rats were selected, aging 4 and half months and weighing 230±15 g.METHODS: The experiment was conducted in the Department of Pharmacology of Guangdong Medical College from May to November 2002.These rats were randomly divided into 3 groups: pseudo-operation group, ovariectomy group and compound norethisterone group, each containing 8 rats. For the former two groups, ethanol solution (volume fraction=0.056), at a dose of 5 mL/(kg.d), was administered by gavage. While for compound norethisterone group, 60μg/(kg·d) norethisterone and 3.5μg/(kg·d) EE were given by gavage (according to the dosage for human, which was 20-35 μg EE combined with norethisterone). Duration of treatment was 90 days for all the animals. Then their tibias were removed. Employing a fullyautomatic imaging analysis system, osteoclasts and the relevant dynamic and static parameters reflecting secondary trabeculaes formation region in proximal tibias were measured. Respectively, the humeral samples were removed and employing the palsma emission spectrograph of full-spectrum direct reading, calcium content and hydroxyproline content in bone samples were measured. Meanwhile, urine calcium and hydroxyproline concentrations were examined as well.MAIN OUTCOME MEASURES: ①The trabecular area (Th. Ar), trabecular thickness (TbkTh), trabecular number (Tb.N) and trabecular separation (Tb. Sp) and the changes in static parareters of perimeters of osteoclasts were investigated. Variance in percent labeled perimeter (L. Pm ％), mineral apposition rate (MAR) and bone formation rate (BFR/BV) were also calculated. ②Changes in serum alkaline phosphatase (AKP), calcium and hydroxyproline contents in bone and urine were all measured.RESULTS: All the 24 rats entered the analysis procedure. Compared to pseudo-operation group, for the ovariectomy group, Tb. Ar and Tb.N decreased, Tb. Sp increased and osteoclast perimeter significantly increased (P＜0.01). Addtionally, the bone formation markers increased apparently with an increase in L. Pm ％ and MAR (P＜0.05) and a significant increase in BFR/BV (P＜0.01). Compared with the ovariectomy group, for the compound norethisterone group,the bone mass and the Tb.N increased, marked by an increase of 82％ in Tb. Ar and an increase of 83％ in Tb.N (P＜0.05), and the Tb.Sp decreased, marked by a decrease of 51％ (P＜0.05). Meanwhile, there was a decrease of 52.5％ in osteoblast perimeter (P＜0.01), an increase in organic bone matrix and a decrease in urine hydroxyproline (P＜0.05).CONCLUSION: A combination of estrogen and progestin has a protective synergistic effect on ovariectomized rats with osteoporosis, and it is capable of increasing the organic bone matrix without significant inhibitory effects on bone formation. The experimental dosage of the compound was calculated according to the clinical dosage, 20-35 μg estrogen combined with a progestin, which will yield optimal protective effects on bone sometimes.

Correlative changes of histomorphometry and biomechanical property of lumbar vertebrae body in ovariectomized rats / 生物医学工程学杂志

This paper presents the correlative changes of cancellous bone histomorphometry, biomechanical property and bone mineral contents of lumbar vertebrae in ovariectomized rats. Forty eight 10.5-month-old female SD rats were randomized to six groups: 1) Basal: at 10.5 mon. of age; 2) sham-1: sham-operated at 13 mon. of age; 3) OVX-1: ovariectomized at 13 mon. of age; 4) sham-2: sham-operated at 16 mon. of age; 5) OVX-2: ovariectomized at 16 mon. of age; and 6) OVX-2 + EE: ovariectomized and treated with 17 alpha-ethynylestradiol (100 micrograms.kg-1.d-1) starting at 13 mon. of age for 12 weeks. Double in vivo fluorochrome labeling was administered to all rats. At the end-point of study, the undecalcified longitudinal fourth lumbar vertebra (LV4) sections were cut and stained with Goldner's Trichrome for bone histomorphometric analyses. The mechanical properties of the compact fifth lumbar vertebra body (LV5) were measured with compression test, the LV5 bodies then were dried constantly with high temperature and digested with acid for testing of bone mineral content. A trend in trabecular bone volume (BV/TV), trabecular thickness (Tb.Th) of LV4, bone break load, break stress, elastic moduli and bone calcium content of LV5 to increase with the increase of age was revealed in the intact rats, but the highest value was at 13 mon. of age. A positive correlation was seen between histomophormetry and bone bio-mechanical properties in the intact rats. Compared with the data from the sham-operated rats, the BV/TV and Tb. Th significantly decreased and bone turnover rate significantly increased in OVX rats at 10 weeks, and the above parameters further decreased 22 weeks post OVX. Correspondingly, the bone break load, break stress, elastic moduli and bone calcium content significantly decreased with the increase of time post OVX was compared with those of sham-operated rats. A positive correlation between histomophormetry and bone biomechanical properties was also observed in OVX rats (r = 0.399, P = 0.018). Bone histomorphometry is good for predicting early physiological and pathological changes and bone biomechanical properties are good parameters for predicting corresponding function of bone such as fracture. Both of them are useful and needful in the animal studies on osteoporosis and related pharmacodynamic observations on new drugs.

Effects of ovariectomized on different parts of bones in rats / 中国临床药理学与治疗学

AIM: To observe the changes of different parts of bones in rats 90 days after ovariectomized (OVX). METHODS: Twenty 4.5 month old virgin female Sprague Dawley rats were randomly divided into 2 groups: sham group and OVX group. Rats in sham group were sham operated, while rats in OVX group were bilaterally OVX. Rats in two groups were treated with 5 ml?kg -1 ?d -1 ethanol for 90 days. Parameters of the proximal tibial metaphysis (PTM), tibial shaft (Tx) and the fifth lumbar vertebral body (LVB) were analyzed by bone histomorphometry method. RESULTS: %Tb.Ar and Tb.N were decreased by 80.5 % and 76.1 %, Tb.Sp and Oc.N were increased by 468.0 % and 356.6 %, and MAR and BFR/BV were increased by 43.9 % and 95.9 %, respectively, in PTM. The changes in LVB were not remarkable as those in PTM. %Tb.Ar and Tb.Th were decreased by 35.0 % and 31.0 %, while Oc.N and BFR/BV were increased by 106.9 % and 126.8 %, respectively. The cortical bone and marrow areas of tibial shaft did not change in Tx, but bone formation parameters (%P LPm, %E LPm) were increased. CONCLUSION: After OVX for 90 days, high bone turnover osteopenia model is duplicated successfully in rats. Different parts of the bones have different reaction to ovariectomization in rats.

Analysis of T-type correlation degree between bone mineral contents and histomorphometry of lumbar vertebrae body in ovariectomized rats / 中国临床药理学与治疗学

AIM: To study the correlative changes of bone mineral contents and histomorphometry of lumbar vertebrae in ovariectomized rats by analysis of T-type correlation degree. METHODS: Forty 10.5-month-old virgin female Sprague-Dawley rats were randomly divided into five groups: (1) Basal: at 10.5 mon of age; (2) sham-1:sham-operated at 13 mon of age; (3) OVX-1: ovariectomized at 13 mon of age; (4) sham-2: sham-operated at 16 mon of age; (5) OVX-2:ovariectomized at 16 mon of age. After ovariectomy, all rats were treated orally with NS at 5 ml?kg -1?d -1. At the end-point of study, the undecalcified longitudinal fourth lumbar vertebra (LV4) sections were cut and stained with Goldner's Trichrome for bone histomorphometric analyses. The fifth lumbar vertebra (LV5) was dried with temperature and digested with acid for testing of bone mineral content. Then the effects of bone mineral contents on bone histomorphometry were assessed by analyzing the T-type correlation degree in the Grey system. RESULTS: All degrees of correlation between bone mineral contents and static histomorphometric parameters (trabecular bone volume) (BV/TV), trabecular thickness (Tb.Th) were positive, but for dynamic histomorphometric parameter (BFR/BV), the correlation degrees were negative. The effect of contents of calcium (Ca) and phosphorus (P) on histomorphometric parameters of lumbar vertebrae was much greater than that of the other bone mineral contents. CONCLUSIONS: Analysis of T-type correlation degree can evaluate the correlative changes of bone mineral contents and histomorphometry of lumbar vertebrae in ovariectomized objectively and fairly.

Effects of epimedium pubescens flavonoids on skeleton of ovariectomized rats / 中国临床药理学与治疗学

AIM: To study the effects of epimedium pubescens flavonoids (EF) on the skeleton in ovariectomized rats. METHODS: Forty 4.5-month-old Sprague-Dawley female rats were randomly divided into 4 groups. Rats in sham group were sham-operated and treated by daily oral gavage with vehicle. Rats in other three groups were bilaterally ovariectomized (OVX) and treated by either daily oral gavage with vehicle, or diethylstilbestrol (DES) at 22.5 ?g?kg~ -1?d~ -1, or EF at 300 mg?kg~ -1?d~ -1 for 90 days. Bone histomorphometric analysis of the proximal tibial metaphysis (PTM), fifth lumbar vertebrae (LV5) and tibial shaft (Tx) was performed in undecalcified sections. The left femur was collected to determine bone weight, contents of calcium (Ca) , phosphorus (P ) and hydroxyproline. The uterine weight and the uterine luminal epithelial thickness (ULET) were determined. RESULTS: A significant increase in contents of Ca and P of femur was found in EF group. A tendency of increase was found in %Tb.Ar of PTM, but no significant change was found in bone histomorphometric parameters of LV5 and Tx in EF group. EF had no effect on uterine weight and ULET. CONCLUSION: EF can prevent OVX-induced bone mineral loss of femur, but does not prevent bone loss of PTM and LV5.

Grey correlative analysis of bone volume and factors in ovariectomized rats / 中国临床药理学与治疗学

AIM: To study the factors that affect bone volume in ovariectomized rats. METHODS: The bone volume and factors were analyzed by the grey system theory method in ovariectomized rats. RESULTS: Serum estradiol was the most important factor for the bone volume, followed by the bone contents of calcium, phosphorus and magnesium. Serum calcium, phosphorus and the bone contents of hydroxyproline were the less important factors for the bone volume. CONCLUSION: Serum estradiol and bone contents of calcium are the most important factors that affect bone volume in ovariectomized rats.KEY WOLDS grey correlative analysis; bone volume; factors; ovariectomized rats; osteoporosis

Combination of norethisterone with ethinylestradiol showed synergetic effect on ovariectomy induced osteopeniain rats / 中国药理学通报

AIM To determine whether norethisterone in combination with ethinylestradiol can completely prevent bone loss in ovariectomized rats. METHODS Twenty-four Sprague-Dawley female rats at the age of 4.5 months were sham-operated and treated orally with vehicle, or ovariectomized (OVX) and treated orally with either vehicle or combined norethisterone (norethisterone at 60 ?g?kg -1?d -1 and ethinylestradiol at 3.5 ?g?kg -1?d -1) for 90 days. Double in vivo fluorochrome labeling was administrated. The undecalcified longitudinal proximal tibial metaphyseal sections were used for the bone histomorphometric analysis. The humerus and urine calcium contents were assayed by the method of atomic absorption spectrometry. The humerus and urine hydroxyproline contents were assayed by colorimetry. Blood serum alkaline phosphatase (ALP) were tested by BECKMAN auto bio-chemistry analyzer. RESULTS After 90 days post OVX the cancellous bone mass and showed high bone turnover indices. Bone hydroxyproline contents were lost markedly. The ALP activity increased. The urine hydroxyproline contents increased significantly. The combined norethisterone treated group prevented bone lost when compared with OVX. The combined norethisterone were shown to inhibit osteoclasts surface and decrese bone turnover rate. The combined norethisterone can increase hydroxyproline contents and reduce urine hydroxyproline contents significantly from OVX group. CONCLUSION Combining norethisterone can prevent OVX-induced cancellous bone loss in rats.

Effects of prostaglandin on osteoporosis in ovariectomized rats / 中国药理学通报

Aim To investigate the preventive effects of misoprostol on osteoporosis in aged ovariectomized(OVX)rats.Methods Female,10-month-old SD rats were ovariectomized(OVX)and,2 months later,were treated with misoprostol or controls for 2 months.The static and dynamic parameters in trabecular bone of the forth lumbar vertebrae(LV4)were examined with histomorphometrical analyses;the fifth lumbar vertebrae(LV5)was used to perform the compression test.Results Compared with the data from the sham-operated rats,the percent trabecular area and elastic modulus significantly decreased in OVX rats.Correspondingly,the bone break load and break stress decreased of post OVX was compared with those of sham-operated rats.Misoprostol increased the percent trabecular area by 21.6% compared with OVX rats,but it couldn't meet the statistical significance.Misoprostol enhanced the break load and elastic modulus compared with OVX rats.Conclusion Misoprostol can improve biomechanics of bone in ovariectomized osteoporosis rats.

Progress in animal models of experimental osteoporsis / 中国临床药理学与治疗学

Animals for model of osteoporosis involve rat, mouse, rabbit, beagle dog, minipig, sheep, etc. The types of model include aged related model, ovariectomized model, orchietomied model, drug treated model, abolition degeneration model, and dietary bone loss. The rat with ovariectomized model is used widely. Biochemical determination, bone mineral density measurement, bone histomorphorphormetry and bone biomechanics are used to judge the formation of experimental osteoporosis.

Diuresis and Removing Urinary Calculus Action of Jieshitong Tablet / 中成药

Objective: Diuresis and removing urinary calculus action of Jieshitong tablet were studied. Methods: Metabolic cage method of rats was adopted to be engaged in diuresis experiment; glycol and ammonium chloride had been given orally for 30 days to form concretion model so that a removing urinary calculus trial could be done. Results: High and low doses of Jeshitong tablet both had significant diuresis effect, total urinary output in 3h of drug group increased about 50% than that of control group. Concretio formative rates of Jeshitong tablet group significantly decresed; the stronger removing urinary calculus action, the higher Jieshitong concentraction was. Conclusion: Jieshitong tablet had diuresis and removing urinary calculus action.

Effects of different doses of cyclophosphamide on bone pharmacology in male rats / 中国药理学通报

AIM To determine the effects of different dose of cyclophosphamide (CP) on the contents of bone calcium and hydroxyproline and the weight of immune organ thymus. METHOD CP at dose of 1 5, 4 5 and 12 5mg?kg -1 were given to the male rats orally everyday for 15 days respectively. At the endpoint the right femoral was dried constantly and weighed (w), the calcium content (Ca.C) was assayed by the method of Atomic Absorption Spectrometry and the femoral hydroxyproline content (Hp.C) were assayed by Colorimetry. The leukocyte, thymus, liver and spleen were tested and weighed separately. RESULTS CP induced inhibiting effect on body weight, leukocyte and thymus in a dose dependent when compared with the vehicle control. Three doses of CP significantly decreased Ca.C and Ca.C/w of femoral ( P0 05). CONCLUSION CP stimulated bone calcium loss, induced shrinkage of thymus. The proper dose at 4 5mg?kg -1 of CP reduced both of Hp.C and Ca.C in bone leading to osteoporosis which is related to the decrease of bone mineral and bone matrix.