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Objective To establish the GIOP model and extract BMSCs from the rat model.We aim to in-vesitigatethe effect ofrhPTH(1-34)for inhibiting β-catenin ubiquitination when combining with Micro-CT and bio-logical technology.We also investigate the influence of rhPTH(1-34)on the GIOP.Methods Female SPF emale rats wererandomly divided into normal control group,methylprednisolone group(model group),methylpredniso-lone+saline group(blankcontrol group)and methylprednisolone+rhPTH(1-34)group(test group). The proximal femoral cancellous bone was examined by Micro-CTand histopathological Staining. The expression of Wnt10b and β-catenin protein were detected. By comparing with inducedBMP-2,BMSCs were treated withrhPTH(1-34)and stained with ALP and alizarin red.Results(1)In Micro-CT,BV/TV,Tb.Th and Tb/N decreased,whereas Tb/sp increased in the test group comparedwith model group(P<0.05).ROI three-dimensional reconstruction of trabecu-lar bone in test group showed local bone repair;(2)Wnt10b and β-cateninexpression increased in the test group compared with the model model(P<0.05),indicating that rhPTH(1-34)can enhance the transcriptional activity of β-catenin(P<0.05)and promote the expression of Wnt10b andβ-catenin(P<0.05).Conclusion The inter-vention with rhPTH(1-34)can prevent GIOP by regulating the Wnt/β-catenin signaling pathway and inhibiting GIOP progress,which can improve the microstructure of bone.
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Objective To investigate the effect of Jinghou Zengzhi Recipe(JHZZR)on Fas apoptosis pathway in ovarian granulosa cells of rats with controlled ovarian hyperstimulation(COH)and to investigate the possible mechanism of the effect of JHZZR on improving oocyte quality. Methods To establish the model of COH,30 rats were randomly divided into the blank group,the positive group and the TCM group.The expression of Fas,Fasl,Caspase8,Caspase3 in ovarian was detected by qPCR andWestern Blot assay,respectively. Results The expression of Fas,Fasl,Caspase8,Caspase3 was lowered in the blank group and the TCM group. Conclusion JHZZR can inhibit Fas signalling,suggesting that Fas pathway may be one of the mechanisms under-lying that JHZZR improves oocyte quality.
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Objective To study the effect of Shenlingbaizhu Pills on hyperlipidemia and gut microbiota in guinea pigs, providing data support for the treatment of hyperlipidemia from the " spleen" . Methods Guinea pigs were randomly divided into control group, model group, ezetimibe group, Shenlingbaizhu Pills high and low dose groups, with 12 guinea pigs for each group. Guinea pig models of hyperlipidemia were established with high fat diet for 8 weeks. Oral administration of the drug began at the fifth week and continued for 4 weeks. The serum lipid levels of guinea pigs were determined after final administration. The pathological changes of liver tissue were observed by HE staining. The mRNA expression levels of ATP binding cassette ( ABC ) A1 in liver, Niemann Pick C1 like 1 ( NPC1L1) and ABCG5/8 in small intestine were detected by realtime PCR. 16S rDNA sequencing was used to detect changes of gut microbiotas in guinea pigs. Results Shenlingbaizhu Pills significantly reduced the serum triglyceride, total cholesterol, and low density lipoprotein-cholesterol levels in guinea pigs with hyperlipidemia ( P＜0. 01 or P＜0.05). The pathological degree of fatty liver was improved obviously by Shenlingbaizhu Pills. Shenlingbaizhu Pills could reduce the mRNA expression level of NPC1L1 of small intestine and increase the level of liver ABCA1 and small intestine ABCG5/8 (P＜0.01 or P＜0.05). At the phylum level, Shenlingbaizhu Pills could reduce the proportion of verrucomicrobia of guinea pigs in model group (P＜0.01); At the genus level, Shenlingbaizhu Pills could increase the proportions of erysipelotrichaceae＿unclassified, ruminococcus＿1, and ileibacterium of guinea pigs in model group (P＜0. 01 or P＜0. 05 ), reduce the proportion of akkermansia and eubacterium＿coprostanoligenes＿group ( P＜0.01). Conclusion Shenlingbaizhu Pills had significantly therapeutic effect on hyperlipidemia, and its specific mechanism was related to the regulation of cholesterol transport and intestinal flora.
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Objective To observe the effects of serum containing Jinghou Zengzhi Recipe (JHZZ) on the expression of growth differentiation factor 9 (GDF9) and Bim in cumulus-oocyte complexes (COCs) and cumulus cells (CCs) of controlled ovarian hyperstimulation (COH) rats,and to investigate its curative effect and mechanism.Methods The rat COH ovarian model was prepared for collecting COCs and CCs,which were respectively co-cultured in vitro with the blank serum of female COH SD rat and serum containing JHZZ,and the each group was divided into 3 subgroups:the blank serum group (blank group),serum containing JHZZ group (control group) and serum containing JHZZ plus GDF9 receptor blocker group (experimental group),total 6 subgroups.COCs and CCs were collected after 24 h.The expression levels of GDF9 and Bim mRNA were detected by real-time quantitative PCR.The GDF9 protein expression levels were detected by Western blot.Results (1) The expression level of GDF9 mRNA in control group COCs was obviously higher than that in the control group and experiment group COCs (P＜0.05);the Bim mRNA expression level in COCs of control group and experiment group was significantly lower than that in COCs of blank group (P＜ 0.05);the GDF9 protein expression level in the control group COCs was significantly higher than that in the blank group COCs (P＜0.05),and also higher than that in the experiment group COCs,but the difference was not statistically significant (P＞0.05).(2)The comparison results of GDF9 mRNA expression level in CCs among 3 groups were consistent with those in COCs;the Bim mRNA expression level in CCs of control group and experimental group was significantly lower than that in the blank group (P＜ 0.05);the Bim mRNA expression level in the control group CCs was significantly lower than that in the experimental group CCs (P＜0.05);the GDF9 protein expression level in the control group CCs was significantly higher than that in the blank group and experimental group CCs (P＜0.05).(3)The GDF9 mRNA expression level in the control group COCs was significantly higher than that in CCs (P＜0.05),and the other inter-group comparisons between COSs and CCs had no statistical difference (P＞0.05).Conclusion Serum containing JHZZ can increase the GDF9 expression level in COCs and CCs,maintain the low expression of Bim in COCs and CCs,inhibit the ovarian cells apoptosis and promote the follicular development;COCs is more conducive to the expression of GDF9 mRNA compared with CCs eliminating oocyte.
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Objective To assess the effects of Jinghou Zengzhi Recipe(JHZZR),a Chinese prescription with the action of tonifying Qi and blood ,on the ovary apoptosis and expression of Bim in ovarian granulosa cells (GCs) of controlled ovarian hyperstimulation(COH) rats , and analyze the possible therapeutic mechanism. Methods A model of COH rats were prepaered and 30 female SD rats were randomly divided into 5 groups , including control group,positive control group,low,medium and high concentration group in six rats in each group. The apoptosis index(AI)in ovarian GCs were detected by TUNEL ,and the expression of Bim by qPCR. Results The AI of ovarian GCs in high and medium concentration group were obviously lower(P 0.05)comparing with control group. The mRNA levels of Bim were lower(P0.05)during the three concentration groups in Bim mRNA. Conclusion JHZZR can inhibit the ovarian GCs apoptosis of COH rats through decreasing the expression of Bim mRNA ,which improve the quality of ovarian follicle.
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Objective To assess the effects of Jinghou Zengzhi Recipe ,a Chinese prescription with the action of tonifying Qi and blood,on the expressions of Bcl-2,Bax and Caspase3 in ovarian granulosa cells of con-trolled ovarian hyperstimulation(COH)mice. Methods A model of COH mice was prepared and 30 female KM mice were divided into blank group,model group and treatment group. The expressions of Bcl-2,Bax and Cas-pase3 were detected by Real-time Quantitative PCR and Western Blot. Results The mRNA and protein levels of Bax and Caspase3 were lower(P<0.05),while the protein levels of Bcl-2 were significantly higher(P<0.05)in the treatment group than those in the model group. There was no significant differences between the treatment group and blank group in the mRNA and proteins. Conclusion Jinghou Zengzhi Recipe can inhibit the ovarian granulo-sa cellular apoptosis of COH mice through increasing the expression of Bcl-2 protein as well as decreasing the ex-pressions of Bax and Caspase3 mRNA and proteins to nearly the natural level,which improves the quality of ovari-an follicle.
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Objective To study whether the effects of bone mineral density by a kidney-tonifying herbal fufang treatment in senile osteoporosis mice (P6) is by the mechanism of improving the expression level of GH mRNA and IGF-1 mRNA. Methods The experimental points four groups as following:SAMR1 mice which feed saline lavage,SAMP6 divid as saline lavage group,subcutaneous injection of rhGH group and a kidney-tonifying herbal fufang treatment group. All intervention is one time everyday. After 3 months and 6 months intervention,we measure the BMD and the expression level of the GH mRNA and of IGF-1 mRNA. Results After 3 months intervention,the BMD of R1 group and the Kidney group were higher than the P6 blank group;but there is no difference in BMD between RhGH group and the P6 blank group. The effect of GH mRNA and IGF-1 mRNA expression levels:the R1 group,rhGH and kidney group were higher than the P6 blank group. After six months intervention,the BMD of the rhGH group and kidney group are higher than the P6 blank group. GH mRNA and IGF-1 mRNA expression levels:GH group and kidney group are higher than the P6 blank group. The expression level of GH mRNA and IGF-1 mRNA in four groups has positive correlation. After six months intervention ,we found the positive correlation between the expression level of GH mRNA and IGF-1 mRNA and each part of the whole body BMD. Conclusion A kidney-tonifying herbal fufang can improve the bone mineral density of P6,and its mechanism may be related to improve expression level of GH mRNA and IGF-1 mRNA.
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Objective To investigate the clinical effect of the Yiqixue Buganshen Recipe(YBR)on the level of serum VEGF in high-risk patients with OHSS. Method 298 infertile women with high-risk OHSS in the reproductive centre of our hospital from June to November ,2016 were enrolled into the traditional Chinese medicine (TCM) group (n = 148) and the control group(n = 150). 145 infertile women without high-risk OHSS were enrolled into the normal group. All the patients received the long protocol treatment ,and patients in the TCM group received the YBR treatment. Results Patients have higher quality embryo rate ,lower moderate and more severe OHSS rate in the TCM group compared with the control group(P<0.05). Patients in the normal group have lower serum VEGF compared with patients in the other 2 groups on the all three days. Patients in the TCM group has lower serum VEGF on the day of embryo transfer compared with patients in the control group(P<0.05). Conclusions The increment of the serum VEGF level is closely related to OHSS. The YBR may reduce the rate of moderate and severe OHSS by reducing the serum VEGF level on the day of embryo transfer ,which may prevent the moderate and severe OHSS of the high-risk patients.
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Objective To explore the establishment methods of animal models of adult growth hormone deficiency, and to provide a good model for experimental research and treatment for abnormal bone metabolism caused by growth hor-mone deficiency.Methods The methods of establishment of animal models of adult growth hormone deficiency were re-viewed and evaluated refering to literature.Results There were three methods including spontaneous lack-of, pituitecto-mized and gene knockout can establish animal models of adult growth hormone deficiency.Conclusions Hypophysecto-mized animal models are inexpensive and easy to create, but not suitable for studying the relationship between growth hor-mone and bone metabolism.Spontaneous lack-of and gene knockout models are specific growth hormone deficiency and of great research significance in exploring the relationship between growth hormone and bone metabolism.
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Objective To observe the effect of Yiqixue Buganshen Recipes(YBR), the compound recipes of Chinese herbal medicine with the actions of replenishing Qi and blood and tonifying liver and kidney, on the endometrial integrin β3 of the mice with embryo implantation dysfunction, and to explore the molecular mechanism of YBR in improving embryo implantation. Methods Ninety mice were divided normal group, model group, and YBR group, 30 mice in each group. YBR group was given Jinghou Zengzhi granules 8mg/kg for promoting post-menstruation proliferation and Cuhuangti granules 8mg/kg for promoting the formation of corpora luteum. At 8∶00 am of pregnant day 4 (Pd4), subcutaneous injection of mifepristone was used for inducing embryo implantation dysfunction in the model group and YBR group. Twenty mice from each group were executed 12 hours after subcutaneous injection of mifepristone, and the uterus was isolated for the detection of mice endometrial integrin β3 mRNA and protein expression with real-time RT-PCR and Western blotting method respectively. Ten mice from each group at Pd4 were executed, and the uterus was isolated and then fixed in 2.5% glutaraldehyde solution for the detection of the formation of pinopode in mice endometrium under scanning electron microscope. Results The detection by real-time PCR and Western blotting method revealed that integrinβ3 mRNA and protein expression levels of the model group were significantly lower than those of the normal group (P<0.05), and YBR group had higher integrin β3 mRNA and protein expression levels than the model group(P<0.05). The formation of pinopodes in the model group was less than the normal group and YBR group, but the number of formed pinopodes in YBR group was similar to that in the normal group. Conclusion YBR can up-regulate the expression of integrin β3 in the endometrium of mice with embryo implantation dysfunction, and have positive regulatory effect on the formation of pinopodes on Pd4.
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Objective To study the mechanism of FTY720 on type Ι diabetic rats. Methods The typeⅠdiabetes rat model was established by feeding male SD rats with high energy urea and injecting into the abdominal cavity with low dose cephalosporins (STZ, 30 mg/kg). In the following, the treated rats were divided into two groups: model group and FTY720 group. Another group of untreated rats was assigned as normal control group. FTY720 group was given 1 mg/kg FTY720, the normal control group and model group was given the equal amount of water. Results The cardiac function of FTY720 group was recovered markeyly. FTY720 activated the expression of vascular endothelial cells S1P1 , diabetes and reduced the expessions of S1P3 and PKCβ Ⅱand it restored the migration ability of diabetes cardiovascular endothelial cell , as well as the abnormally elevated cardiovascular endothelial cells induced by high sugar permeability. Conclusion The S1P1 and S1P3 cut is an important reaction pathway to the complications of diabetes and cardiovascular dysfunction. FTY720 may reduce the damage to core blood vessels caused by diabetes , and pathological angiogenesis which functionally depends on the PKCβⅡ signaling pathways. Therefore, FTY720 may provide a potential therapy for cardiovascular function impaired by diabetes.
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Objective To investigate the concentration of 17β-estradiol in bone marrow stem cells (BMSCs) and different tissues of rats, and study the function of estradiol produced in extragonadal sites prelimi-narily. Methods (1) The concentration of 17β-estradiol in lysate of BMSCs and in supernatant cell culture media were detected by Elisa method after rats BMSCs were serum-free cultured in 0 h , 24 h and 48 h respective ly . (2) The tissues of organs were grinded and broken under ultrasonic wave , then washed and weighed. The con-centration of 17β-estradiol in different tissues was detected by Elisa method. Results Compared with the concen-tration of 17β-estradiol at 24 h, the concentration of 17β-estradiol at 48h significantly increased (P 0.05). Conclusions (1) 17β-estradiol can be secreted by BMSCs , and the concentration is proportional to the time to some degree. But whether the extragonadal estrogen can function locally is still unclear. (2) The concentration of 17β-estradiol is non- gonad dependent but whether it is secreted locally remains to be elucidated. The non- gonad estrogen could be the estrogen source of bone metabolism in order to sustain bone health after menopause.
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BACKGROUND:Studies have shown that genetical y modified Schwann cel s can survive for a longer time in vivo, and promote nerve regeneration and functional recovery. OBJECTIVE:To transfect human telomerase reverse transcriptase (hTERT) gene into rat Schwann cel s cultured in vitro via PLXSN vector, and to detect the telomerase activity and biological characteristics of Schwann cel s. METHODS:Schwann cel s from Wistar rats were cultured in vitro and transfected by PLXSN vector with (hTERT group) or without hTERT (empty vector group). Normal Schwann cel s were selected as control group. RT-PCR and western blot methods were used to detect the hTERT protein and mRNA levels in Schwann cel s, and flow cytometry was used to measure the cel cycle distribution. Cel growth was observed by cel growth curve and MTT colorimetric method. RESULTS AND CONCLUSION:At 48 hours after transfection, the mRNA and protein expressions of hTERT were remarkably seen in Schwann cel s. Compared with the control and empty vector groups, the cel s grew faster, the number of cel s at G 0/G 1 Schwann cel s cultured in vitro. phase was reduced, but the number of S phase cel s was increased in the hTERT group (P<0.05). These findings indicate that PLXSN vector-mediated hTERT transfection of Schwann cel s can significantly improve the activity of telomerase in Schwann cel s as wel as promote the proliferation of Schwann cells cultured in vitro.
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BACKGROUND:Estrogen signaling pathway for interaction between aromatase and estrogen-related receptor may exist in bone marrow mesenchymal stem cel s, which is used for regulating biological activity of bone marrow mesenchymal stem cel s. OBJECTIVE:To observe the expression of aromatase and estrogen-related receptors in adult bone marrow mesenchymal stem cel s during osteogenic differentiation. METHODS:Bone marrow mesenchymal stem cel s were respectively cultured in low-glucose DMEM medium (control group) and osteogenic induction medium (induction group). Cel proliferation and calcium deposition were determined by MTT assay and alizarin red staining, respectively. The expression of aromatase, estrogen receptorα, estrogen receptorβ, and estrogen-related receptorαduring osteogenic differentiation were determined by real-time PCR and western blot analysis. Estradiol levels in supernatants and lysates were detected by ELISA method. RESULTS AND CONCLUSION:In the induction group, the proliferation ability of bone marrow mesenchymal stem cel s was the strongest at 72 hours of culture;while there were a great amount of calcium nodules formed at 21 days of culture. Results from PCR and western blot assay showed that the expression of aromatase and estrogen receptorαwas improved in the induction group, but the expression of estrogen-related receptorαwas inhibited. There was no difference in the expression of estrogen receptorβbetween the two groups. ELISA results indicated that the level of estradiol in the supernatant of induction group was the highest. These findings indicate that aromatase, estrogen receptorα, estrogen receptorβand estrogen-related receptorαare al involved in osteogenesis of bone marrow mesenchymal stem cel s. Moreover, estradiol can be synthesized and secreted in bone marrow mesenchymal stem cel s, and most likely, promote the osteogenic differentiation of bone marrow mesenchymal stem cel s by related receptor pathway.
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Objective To investigate the relationship between the serum SRC-3 levels and the bone loss severity in postmenopausal women. Methods Fifty-eight PMW with osteopenia or osteoporosis and nineteen healthy PMW were enrolled in this study from June 2012 to September 2013. BMD at the lumbar spine and femoral neck were observed by DXA Lunar Prodigy Vision. The levels of serum SRC-3 were detected by ELISA. The diagnosis value was evaluated by the ROC curves analysis. Results The levels of serum SRC-3 were significant higher in the normal group than those in the osteopenia or the osteoporosis groups (P<0.001 for both), no statistical significance was found between the osteopenia and the osteoporosis group(P=0.056). The levels of serum SRC-3 were negatively correlated with the BMD diagnosis grading (r=-0.543, P < 0.001). By using the ROC curve analysis, the serum level of SRC-3 for PMW with osteoporosis and osteopenia were found to be 0.297 ng/mL and 0.347 ng/mL, respectively. The levels of serum SRC-3 were positively associated with BMI (r=0.395, P<0.001) and LS-BMD (r=0.503,P<0.001) in the postmenopausal women. Conclusion SRC-3 might be an useful index to reflect the severity of lumbar spine bone loss.
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Objective:To investigate the effect of staphylococcus aureus on expressions of inducible nitric oxide synthase (iNOS) and interleukin-1 beta (IL-1β) in THP-1 monocytes under high glucose concentrations. Methods:THP- 1 monocytes were incubated at different surroundings with 2×2 factorial design. There were 4 experimental groups in the study, which were analyzed by univariate analysis of variance. The total RNA was distilled. The expressions of iNOS and IL-1β were examined by SQ-RT-PCR. Results:The expressions of iNOS and IL-1β were affected by high glucose concentration, bacteria and both of them together in THP-1 monocytes. The expressions of iNOS and IL-1β were weakened in high glucose concentration compared with that of control(P < 0.01). The expressions of iNOS and IL-1β were weakened in the high glucose concentration and bacterial infection compared with that of the high glucose concentration only(P < 0.01). The expressions of iNOS and IL-1β increased in bacterial infection compared with that of control(P < 0.01). The expression of IL-1β increased in high glucose concentration and bacterial infection compared with that of the high glucose concentration only(P < 0.01) but the expression of iNOS was not distinct. Conclusion:When staphylococcus aureus infected monocytes, the expressions of iNOS and IL-1β were weakened in high glucose concentration, which suggested that the depressed function of immunocyte was related with high glucose concentration.
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Objective To investigate the possibility of immune tole rance induced by bone marrow-derived mesenchymal stem cells in allogeneic organ transplantation. Methods Allogeneic bone marrow-derived mesenchymal stem cells and syngeneic bone marrow cells were cotransplanted to lethally irradiated female C57BL/6 recipient mice. FACS was used to analyze the chimerism 150 days later. Mixed lymphocyte reaction and ConA induced proliferation test were performed to evaluate proliferative activity of mice spleen cells in cell-transplanted group. Skin transplantation test was done to observe immune response of cell-transplanted group mice against organ graft from donor mice. Results About 5 97% donor T cells were detected in splenocytes of cell-transplanted group mice. MLR showed that mean SI of cell-transplanted group mice was 1.79, and that of untreated group mice was 7 28. ConA induced proliferation test showed that mean SI of cell-transplanted group mice was 31 92; and that of untreated group mice was 34 99. Mean survival time of donor-derived skin graft in cell-transplanted group mice was more than 90 days; and that in untreated group mice was 8 days. Conclusion Our results showed for the first time that induction of stable mixed hematopoietic chimerism after allogeneic bone marrow derived mesenchymal stem cells transplantation lead to stable donor-specific tolerance in allogeneic host and skin graft survival from donor mice.
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Objective To explore the influences of Bushen Zhuanggu Granule (BZG)on contents of sexual hormones and metabolism of calcium and phosphorous in early menopausal women.Methods One hundred and Twenty-two early menopausal women were allocated to two groups:Group A was treated with BZG and Group B with nylestriol.Serum contents of sexual hormones and indexes of calcium and phosphorous metabolism were observed before and after treatment.Results After six months of treatment,the contents of serum estradiol(E 2)and alkaline phosphatase(ALP)were increased and the ratio of urine calcium and creatinine(Ca/Cr)decreased(P 0.05).The contents of follicle stimulating hormone(FSH)and luternizing hormone(LH)were increased in Group A as compared with Group B(P