Clinical Observation on the Effect of Postmenstrual Proliferative Prescription Through Replenishing Qiand Blood on Ovulation Induction in Patients with Polycystic Ovarian Syndrome / 广州中医药大学学报

Objective To observe the effects of Postmenstrual Proliferative Prescription(mainly composed of Codonopsis Radix,Atractylodis Macrocephalae Rhizoma,Poria,Rehmanniae Radix Praeparata,Paeoniae Radix Alba,Angelicae Sinensis Radix,Chuanxiong Rhizoma,Cervi Cornu Degelatinatum,Corni Fructus,Cuscutae Semen,and Eucommiae Cortex)through replenishing qi and blood on the ovulation rate and pregnancy rate in patients with ovulatory dysfunction infertility caused by polycystic ovarian syndrome(PCOS)during ovulation-induction treatment,and to explore the therapeutic effects and possible therapeutic mechanism.Methods Sixty patients with ovulatory dysfunction infertility due to PCOS were randomly divided into a treatment group and a control group,with 30 patients in each group.The control group was given Clomifene Citrate Capsules to promote ovulation,and the treatment group was given Postmenstrual Proliferative Prescription on the basis of the ovulation-induction program of the control group starting from the fifth day of menstruation or progesterone withdrawal bleeding.The two groups were treated for one menstrual cycle as a course of treatment.The changes in the serum sex hormones of estradiol(E2),follicle-stimulating hormone(FSH),luteinizing hormone(LH),and progesterone(P)on the 2nd to 5th day of menstruation,as well as the changes in serum growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)levels on the 2nd to 5th day of menstruation and on the day of human chorionic gonadotropin(HCG)injection were observed in the two groups.Moreover,the ovulation rate,pregnancy rate and clinical efficacy of the patients in the two groups were analyzed.Results(1)No statistically significant differences in serum levels of sex hormones E2,FSH,LH and P on the 2nd to 5th day of menstruation were shown between the two groups of patients(P＞0.05).(2)On the 2nd to 5th day of menstruation and on the day of HCG injection,there were no significant differences in the serum GDF9 and BMP15 levels between the two groups(P＞0.05).(3)The ovulation rate and pregnancy rate in the treatment group were 93.33%(28/30)and 26.67%(8/30)respectively,which were significantly higher than 70.00%(21/30)and 13.33%(4/30)in the control group.And the differences tested by chi-square test were statistically significant between the two groups(P＜0.05).(4)The total effective rate of the treatment group was 93.33%(28/30),and that of the control group was 70.00%(21/30).The intergroup comparison(tested by rank sum test)showed that the therapeutic efficacy of the treatment group was significantly superior to that of the control group,and the difference was statistically significant(P＜0.05).Conclusion Postmenstrual Proliferative Prescription through replenishing qi and blood can improve the ovulation rate and pregnancy rate during ovulation-induction treatment in the patients with ovulatory dysfunction infertility due to PCOS.It is indicated that Postmenstrual Proliferative Prescription can enhance the quality of the oocytes and the potential of embryo implantation during the ovulation-induction treatment.

Jiegu Qili Capsules Promotes Fracture Healing of Radius in Rats by Activating the BMP/Smad Signaling Pathway / 广州中医药大学学报

Objective To investigate the improvement effect of Jiegu Qili Capsules on the fracture healing of radius in rats by activating the bone morphogenetic protein(BMP)/Smad signaling pathway.Methods(1)A rat model of radius fracture was constructed,and the serum levels of alkaline phosphatase(ALP),calcium(Ca)and phosphorus were detected,and the pathological changes in the fracture gap was observed.(2)Human osteosarcoma SaOS-2 cells were used to measure ALP activity and mineralization level.The quantitative real-time polymerase chain reactionn(qRT-PCR)method was used to detect the cellular osteogenesis-related genes ALP,collagen I(COL-I),ornithine transcarbamylase(OTC),Osterix,osteoblastin(OPN),Runt-related transcription factor 2(RUNX2)and BMP2.The expression of key proteins in BMP/Smad signal pathway was detected by Western Blot.Results Jiegu Qili Capsules effectively promoted fracture healing of radius in rats,enhance ALP activity,increase Ca and P levels Jiegu Qili Capsules stimulate the formation of mineralized nodules in SaOS-2 cells in rats.,and promoted the expression levels of COL-I,OTC,Osterix,BMP2 and OPN in SaOS-2 cells in a dose-dependent manner.Jiegu Qili Capsules up-regulated the levels of Smad1/5 phosphorylation of the BMP/Smad signaling pathway in SaOS-2 cells,as well as the levels of BMP2 and RUNX2.Noggin,an inhibitor of the BMP/Smad signaling pathway,inhibited osteogenic differentiation of SaOS-2 cells induced by Jiegu Qili Capsules.Conclusion Jiegu Qili Capsules can promote fracture healing by activating the BMP/Smad signaling pathway to increase the expression of osteogenesis-related genes.

Expression of bone morphogenetic protein 9 in patients with sepsis-associated acute respiratory distress syndrome and its role in early recognition and prognosis prediction of disease / 中华急诊医学杂志

Objective:To observe the expression level of bone morphogenetic protein 9 (bone morphogenetic protein 9,BMP9) in patients with sepsis-associated acute respiratory distress syndrome (acute respiratory distress syndrome,ARDS), and to explore the role of BMP9 in early recognition and prognosis prediction of sepsis-associated ARDS.Methods:From May 2022 to May 2023, total of 56 patients with sepsis-associated ARDS in Shanxi Bethune Hospital were selected as the ARDS group, 49 patients with cardiogenic pulmonary edema as the case control group, and 46 adults who underwent physical examination in the physical examination center of our hospital as the healthy control group.The patients in the ARDS group were followed up for 28 days and divided into survival group ( n = 26) and death group ( n = 30). The expression level of serum BMP9 and its correlation with clinical indicators in each group were analyzed and compared. The risk factors of sepsis-associated ARDS were analyzed by Logistic regression, and the diagnostic efficacy and prognostic value of related indicators were analyzed. Results:The serum level of BMP9 in sepsis-associated ARDS group [1401.14 (856.59,1982.86) ]pg/mL was significantly higher than that in case control group (438.26±128.52) pg/mL and healthy control group (398.96±96.55)pg/mL, the differences were statistically significant ( P<0.01). In addition, BMP9 expression significantly correlated with procalcitonin, lymphocyte count and SOFA score ( P < 0.05, P < 0.01, respectively). Multivariate Logistic regression analysis showed that BMP9 was a high risk factor for the development of sepsis-associated ARDS ( P<0.01). The area under the ROC curve (area under the ROC curve,AUC) of BMP9 to predict the occurrence of sepsis-associated ARDS was 0.930. The specificity was 100.0% and the sensitivity was 80.4%, which was significantly higher than the specificity (89.8%) and sensitivity (67.9%) of the oxygenation index. Follow-up and comparison of BMP9 levels in patients with different prognosis of sepsis-associated ARDS showed that the expression level of BMP9 in the death group was higher than that in the survival group, and the difference was statistically significant ( P < 0.05). The ROC curve of BMP9 in predicting the prognosis of patients with sepsis-associated ARDS. The area under the ROC curve was 0.699, the sensitivity was 43.3%, and the specificity was 100.0%. Conclusions:The expression of BMP9 in sepsis-associated ARDS patients significantly increased, and its high expression was significantly correlated with inflammatory markers such as procalcitonin, lymphocyte count and SOFA score. BMP9 is an independent risk factor in patients with sepsis-associated ARDS, and it is promising as a new biomarker for early identification of sepsis-associated ARDS. However, it do not show a good predictive effect on the prognosis of the disease.

Irisin alleviates palmitic acid-induced osteogenic inhibition in bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Irisin,a myokine isolated from the transmembrane protein FNDC5 by muscle cells during exercise,has the function of inducing the browning of white adipose tissue,but its effect on lipotoxicity-induced osteogenic differentiation and the mechanism is unclear. OBJECTIVE:To investigate the effect of irisin on the osteogenic ability of palmitic acid-induced bone marrow mesenchymal stem cells and the mechanism of action. METHODS:CCK-8 assay was used to detect the effect of different concentrations of palmitic acid on the proliferation of mouse bone marrow mesenchymal stem cells and the effect of irisin on the proliferation of mouse bone marrow mesenchymal stem cells in the presence of palmitic acid.After pretreatment with irisin and palmitic acid for 24 hours,osteogenic differentiation of mouse bone marrow mesenchymal stem cells was induced by alkaline phosphatase staining as well as qRT-PCR was performed to detect the expression of osteogenesis-related genes on day 7 of osteogenic induction culture.The expression of proteins related to the AMPK/BMP2/SMAD signaling pathway was detected by western blot assay.Alizarin red staining was conducted on day 21 to detect osteogenic differences. RESULTS AND CONCLUSION:(1)The CCK-8 assay results suggested that the amplification of bone marrow mesenchymal stem cells was inversely proportional to the concentration of palmitic acid,but at 0.02 mmol/L concentration,palmitic acid had no significant effect on the amplification of bone marrow mesenchymal stem cells,and irisin did not affect the proliferation of bone marrow mesenchymal stem cells when its mass concentration was in the range of 0.1-20 μg/L.(2)Alkaline phosphatase staining and alizarin red staining showed that palmitic acid inhibited the osteogenic differentiation ability of bone marrow mesenchymal stem cells.Irisin improved palmitic acid-induced osteogenic inhibition of bone marrow mesenchymal stem cells.qRT-PCR results showed that palmitic acid could cause the downregulation of osteogenic-related genes,and irisin could inhibit this trend.(3)Western blot assay results showed that compared with the palmitic acid intervention group,irisin treatment enhanced AMPK/BMP2/SMAD signal transduction in bone marrow mesenchymal stem cells.It is found that irisin can improve the osteogenic differentiation ability of bone marrow mesenchymal stem cells pretreated with palmitic acid,and proposed that the specific mechanism might be mediated by AMPK/BMP/SMAD signaling pathway.

Bone morphogenetic protein/Smad signaling pathway involved in chondrodysplasia in the ankle of a rat congenital clubfoot model / 中国组织工程研究

BACKGROUND:Congenital clubfoot mainly manifests as abnormal bone itself and abnormal cartilage development.The bone morphogenetic protein(BMP)/drosophila mothers against decapentaplegic protein(Smad)signaling pathway can direct the development of bone and cartilage during embryonic period,but its role in the field of clubfoot etiology has not been confirmed in animal experiments. OBJECTIVE:To explore the mechanism by which the BMP/Smad signaling pathway is involved in the regulation of foot and ankle chondroplasia in a rat congenital clubfoot model. METHODS:Sprague-Dawley rats at 10 days of gestation with the same growth condition were randomly divided into experimental and control groups.The experimental group was intragastrically given 135 mg/kg retinoic acid to make the clubfoot model,while the control group was given the same amount of vegetable oil.The fetal rats were taken out after 21 days of gestation by cesarean section.In the experimental group,the 27 of 41 fetal rats had clubfoot,with a deformity rate of 65.9%;in the control group,no clubfoot was observed in all the 36 fetuses.The ankles tissues of the rats were collected for hematoxylin-eosin staining.Western blot assay,RT-qPCR and immunohistochemistry were performed to detect the expression levels of Smad5 and P-Smad5,the core proteins of the BMP/Smad signaling pathway,as well as SP7 and Sox9,the downstream proteins of the BMP/Smad signaling pathway. RESULTS AND CONCLUSION:Compared with the control group,hematoxylin-eosin staining showed that the cartilage matrix in the foot and ankle tissues increased and the gap between the bones increased in the experimental group.Immunohistochemical findings showed that the expression levels of Smad5 and SP7 decreased in the experimental group,while the mRNA expression of Sox9 increased.RT-qPCR results showed that the mRNA expression of Smad5 and SP7 decreased,while the mRNA expression of Sox9 increased in the foot and ankle tissues of rats in the experimental group.Western blot results showed that P-Smad5/Smad5 expression and SP7 expression were decreased and Sox9 expression was increased in the ankle of rats in the experimental group.To conclude,the occurrence of cartilage abnormalities in the foot and ankle of the rat model of congenital clubfoot is associated with impaired transmission of the BMP/Smad signaling pathway.

Mechanism of Youguiwan Regulating BMP-2/Smad Signaling Pathway to Promote Bone Formation in Postmenopausal Rats with Osteoporosis / 中国实验方剂学杂志

ObjectiveTo observe the effects of Youguiwan on bone metabolism and bone morphogenetic protein-2 （BMP-2）/Smad signaling pathway in ovaries-removed rats with osteoporosis and study the mechanism of Youguiwan in the prevention and treatment of osteoporosis. MethodA postmenopausal rat model of osteoporosis was prepared by bilateral ovariectomy. The 40 female SD rats were randomly divided into five groups， including sham operation group， model group， alendronate sodium group （0.1 mg·kg-1）， and high-dose and low-dose （5.36 and 2.68 g·kg-1） groups of Youguiwan. The drug was given seven days after modeling， once a day for 12 weeks. After the treatment， the changes in femur tissue structure were observed by micro-CT， including bone mineral density （BMD）， bone volume/total volume （BV/TV）， trabecular number （Tb.N）， trabecular thickness （Tb.Th）， bone surface/bone volume （BS/BV）， and trabecular separation （Tb.Sp）. Ossification was observed by saffrane-solid green staining， and serum levels of bone metabolism markers， including bone alkaline phosphatase （BALP）， osteocalcin （BGP）， type Ⅰ procollagen amino terminal propeptide （PINP）， and tartrate-resistant acid phosphatase 5b （TRACP-5b）， were determined by enzyme-linked immunosorbent assay （ELISA）. The protein and mRNA expression levels of Runx2， BMP-2， and Smad1 in rat femur were detected by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the sham operation group， bone trabecula in the model group was sparse. BMD， BV/TV， Tb.N， and Tb.Th were decreased （P<0.05， P<0.01）. BS/BV （P<0.05） and Tb.Sp were increased. The content of BGP， BALP， PINP， and TRACP-5b in serum was significantly increased （P<0.01）. The mRNA and protein expressions of Runx2， BMP-2， and Smad1 in rat femur were significantly decreased （P<0.05， P<0.01）. Compared with the model group， the number of bone trabeculae in the high-dose and low-dose groups of Youguiwan was increased， and the bone microstructure was improved. BMD， BV/TV， Tb.N， and Tb.Th were increased significantly （P<0.05， P<0.01）， and BS/BV and Tb.Sp were increased. The content of bone metabolic markers decreased （P<0.05， P<0.01）. ConclusionYouguiwan has certain preventive and therapeutic effects on postmenopausal osteoporosis， and its mechanism may be related to promoting bone formation by regulating the BMP-2/Smad signaling pathway.

Osteoprotective Effect and Mechanisms of Taxillus chinensis from Different Hosts on Ovariectomized Osteoporotic Rats / 中国实验方剂学杂志

ObjectiveTo compare the effects of Taxillus chinensis from different hosts with different meridian affinity on bone microstructure and bone metabolism in ovariectomized osteoporotic rats， and investigate its mechanism of action. MethodEighty-eight specific-pathogen-free （SPF）-grade female Sprague-Dawley （SD） rats were selected and randomly divided into 11 groups： sham-operated group， model group， low-， medium- and high-dose groups of T. chinensis from Morus alba （2.5， 5， and 10 g·kg-1）， low-， medium- and high-dose groups of T. chinensis from Cinnamomum cassia （2.5， 5， and 10 g·kg-1）， and low-， medium- and high-dose groups of T. chinensis from C. burmannii （2.5， 5， and 10 g·kg-1）. After 12 weeks of drug intervention， the rats were examined for proximal femur bone density and bone microstructure using dual-energy X-ray absorptiometry （DXA） and micro-computed tomography （Micro-CT）. Histopathological changes in rat femur were observed by the hematoxylin-eosin staining （HE）. Contents of serum estradiol （E2）， bone Gla protein （BGP）， bone alkaline phosphatase （BALP）， tartrate-resistant acid phosphatase 5b （TRACP-5b） and pre-collagen type Ⅰ amino-terminal protopeptide （PINP） were measured by the enzyme-linked immunosorbent assay （ELISA）. Real-time quantitative polymerase chain reaction （Real-time PCR） was employed to detect the messenger ribonucleic acid （mRNA） expressions of bone morphogenetic protein-2 （BMP-2）， Smad1， Smad9 and recombinant runt-related transcription factor 2 （Runx2） in rat humerus. Western blot was used to detect the protein expressions of BMP-2， p-Smad1/5/9 and Runx2 in rat humerus. ResultCompared with that in the sham-operated group， the femur microstructure of rats in the model group was significantly disrupted， with significant decreases in bone mineral density （BMD） value， bone volume fraction （BV/TV）， trabecular number （Tb.N）， and trabecular thickness （Tb.Th） （P<0.01）， and significant increases in trabecular separation （Tb.Sp） and structure model index （SMI） （P<0.01）. The serum levels of BGP， BALP， TRACP-5b and PINP were significantly increased （P<0.05， P<0.01）， and E2 levels were significantly decreased （P<0.01）. The mRNA expressions of BMP-2， Smad1， Smad9， and Runx2 were significantly decreased in rat humerus （P<0.01）， and the protein expressions of BMP-2， p-Smad1/5/9， and Runx2 were significantly reduced （P<0.01）. Compared with the model group， the administration groups of T. chinensis from different hosts all elevated the BMD， BV/TV， Tb.N， Tb.Th， Tb.Sp， and SMI levels in the femur， improved bone microstructure， increased serum E2 levels （P<0.05， P<0.01）， lowered the levels of serum BGP， BALP， TRACP-5b， and PINP， upregulated the mRNA expression of BMP-2， Smad1， and Runx2 and upregulated the mRNA expression levels of Smad9 （P<0.05， P<0.01）， and upregulated the protein expressions levels of BMP-2， p-Smad1/5/9， and Runx2 （P<0.01）. The best effect was observed in the group of T. chinensis from C. cassia. ConclusionT. chinensis from different hosts improved osteoporosis in ovariectomized rats， with the group of T. chinensis from C. cassia being the most potent among the administered groups， and its treatment of osteoporosis may regulate the balance of bone conversion by regulating BMP/Smad signaling pathway.

BMP7 expression in mammalian cortical radial glial cells increases the length of the neurogenic period

The seat of human intelligence is the human cerebral cortex, which is responsible for our exceptional cognitive abilities. Identifying principles that lead to the development of the large-sized human cerebral cortex will shed light on what makes the human brain and species so special. The remarkable increase in the number of human cortical pyramidal neurons and the size of the human cerebral cortex is mainly because human cortical radial glial cells, primary neural stem cells in the cortex, generate cortical pyramidal neurons for more than 130 days, whereas the same process takes only about 7 days in mice. The molecular mechanisms underlying this difference are largely unknown. Here, we found that bone morphogenic protein 7 (BMP7) is expressed by increasing the number of cortical radial glial cells during mammalian evolution (mouse, ferret, monkey, and human). BMP7 expression in cortical radial glial cells promotes neurogenesis, inhibits gliogenesis, and thereby increases the length of the neurogenic period, whereas Sonic Hedgehog (SHH) signaling promotes cortical gliogenesis. We demonstrate that BMP7 signaling and SHH signaling mutually inhibit each other through regulation of GLI3 repressor formation. We propose that BMP7 drives the evolutionary expansion of the mammalian cortex by increasing the length of the neurogenic period.

Growth Differentiation Factor 5 and Metabolic Diseases / 生物化学与生物物理进展

Growth/differentiation factor-5 (GDF-5) belongs to transforming growth factor-β (TGF-β) family, which is expressed in bone, cartilage, heart, brain, kidney, skeletal muscle and tendon, liver, fat and other organs and tissues as well. GDF-5 binds to receptor BMPR-I/BMPR-II and activates different signaling pathways such as smad1/5/8, PI3K/Akt, p38-MAPK. For a long time, numerous studies have shown that GDF-5 plays an important role in protecting joints. However, researchers have found GDF-5 also plays significant biological functions in other organs. For example, GDF-5 improves cardiac function by reducing oxidative stress and fibrosis in infarcted hearts. GDF-5 can also reduce oxidative stress in the brain and increase the number of neurons in effort to delay the progression of Alzheimer’s disease and Parkinson’s disease. It is a situation, research on GDF-5, at present, mainly focuses on the growth and repair of bone, cartilage and tendons, while there are few reports on its biological effects in other organs. Therefore, this article reviews and summarizes the research progress on GDF-5 and metabolic diseases in recent years in order to provide new insights and theoretical basis for the role of GDF-5 in improving metabolic diseases.

Mutations in the bone morphogenetic protein 15 gene causing defective protein secretion in Cholistani infertile sheep / Mutações no gene da proteína morfogenética óssea 15 causadoras de defeitos na secreção proteica em ovelhas inférteis Cholistani

Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.

Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.

Mutations in the bone morphogenetic protein 15 gene causing defective protein secretion in Cholistani infertile sheep

Abstract Naturally occurring mutations in morphogenetic protein 15 (BMP15) are associated with decreased ovulation rate (OR), litter size (LS), and sterility. It is of a great interest to elucidate BMP15 gene in Cholistani sheep breed to uplift socio-economic status and the knowledge of Cholistani sheep breeding in Southern Punjab, Pakistan. In our study, a total of 50 infertile Cholistani sheep aged between 2-6 years and having no blood relation were screened for BMP15 mutations. For this purpose, a high-quality DNA was extracted from the blood of sheep followed by primer designing, Polymerase Chain Reaction (PCR) amplification, DNA sequencing, and in silico analyses. Out of total 50 samples, 9 samples including case 1 (T3), case 2 (T8), case 3 (T17), case 4 (T22), case 5 (T25), case 6 (T33), case 7 (T40), case 8 (T44), and case 9 (T47) were found positive for a variety of already reported and novel BMP15 mutations. Further in silico analyses of the observed mutations have shown the functional impact of these mutations on different characteristics (molecular weight, theoretical PI, estimated half-life, instability index, sub-cellular localization, and 3D confirmation) of the encoded proteins, possibly altering the normal functionality. In a nutshell, findings of this study have confirmed the possible essential role of the BMP15 mutations in the infertility of the Cholistani sheep.

Resumo Mutações de ocorrência natural na proteína morfogenética 15 (BMP15) estão associadas à diminuição da taxa de ovulação (TO), tamanho da ninhada (TN) e esterilidade. Estudar a BMP15 na raça Cholistani para elevar o status socioeconômico e o conhecimento da criação de ovinos Cholistani no sul de Punjab, Paquistão. Em nosso estudo, 50 ovelhas Cholistani inférteis sem parentesco sanguíneo foram rastreadas para mutações BMP15. Para tanto, um DNA de alta qualidade foi extraído do sangue dessas ovelhas, seguido de concepção do primer, amplificação da reação em cadeia da polimerase (PCR), sequenciamento de DNA e análises in silico. Do total de 50 amostras, 9, incluindo caso 1 (T3), caso 2 (T8), caso 3 (T17), caso 4 (T22), caso 5 (T25), caso 6 (T33), caso 7 (T40), caso 8 (T44) e caso 9 (T47), foram consideradas positivas para uma variedade de mutações BMP15 novas e já relatadas. Mais análises in silico das mutações observadas mostraram o impacto funcional dessas mutações em diferentes características (peso molecular, PI teórico, meia-vida estimada, índice de instabilidade, localização subcelular e confirmação 3D) das proteínas codificadas, possivelmente alterando a funcionalidade normal. Nossos achados confirmaram o possível papel essencial das mutações BMP15 na infertilidade de ovelhas Cholistani.

Comparative study of the effects of intramedullary nail fixation and minimally invasive percutaneous plate internal fixation technique on platelet activation and serum transforming growth factor-β1(TGF-β) 1 and bone morphogenetic protein-2 (BMP-2) in patients with tibial and fibular fracture / 中国骨伤

OBJECTIVE@#To investigate the effect of intramedullary nail fixation (IMN) and minimally invasive percutaneous plate internal fixation (MIPPO) techniques on tibiofibular fractures and their effect on platelet activation and serum transforming growth factor-β1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2).@*METHODS@#Total of 105 patients with tibiofibular fractures from February 2019 to February 2020 were selected and divided into 53 cases in the MIPPO group and 52 cases in the IMN group. There were 29 males and 24 females with an average age of (41.74±6.05) years old in MIPPO group;in IMN group, 31 males and 21 females with an average age of (40.59±5.26) years old. The perioperative surgical indexes, postoperative complications, ankle function recovery at 12 months postoperatively, platelet activation indexes at 3 and 7 days preoperatively and postoperatively, and serum TGF-β1 and BMP-2 levels at 4 and 8 weeks preoperatively and postoperatively were compared between the two groups.@*RESULTS@#The operating time and fracture healing time in the MIPPO group were shorter than those in the IMN group(P<0.05); Compared with the preoperative period, the levels of GMP-140, PAC-1, CD63, and CD61 increased in both groups at 3 and 7 days after surgery, but were lower in the MIPPO group than in the IMN group(P<0.05);the levels of serum TGF-β1 and BMP-2 increased in both groups at 4 and 8 weeks after surgery compared with the preoperative period, and the postoperative complication rate in the MIPPO group was lower than that in the IMN group(P<0.05);the difference was not statistically significant in the excellent rate of ankle function recovery at 12 months follow-up after surgery between two groups(P>0.05).@*CONCLUSION@#Both intramedullary nail fixation and MIPO technique for treatment of tibia and fibula fractures can improve ankle joint function, but the latter has the advantages of short operation time, fast fracture healing, fewer complications, and light platelet activation. Serum TGF-β1, BMP-2 level improves quickly.

Isopsoralen Promotes Osteoblast Proliferation and Differentiation via BMP2/Runx2/Osx Signaling Pathway / 世界科学技术-中医药现代化

Objective The purpose of this study was to examine effects of Isopsoralen on the osteoblast proliferation and differentiation and find its possible molecular mechanisms for anti-osteoporosis.Methods OCT-1 cells were cultured with common methods.While growing well,cells were cultured with 3 doses(10 μg·mL-1,30u μg·mL-1 and 60 μg·mL-1)of Isopsoralen for 48 h,or with purified bone morphogenetic protein 2(BMP2)protein(50 ng·mL-1).We first determined the effect of Isopsoralen on cell proliferation by MTT assay.The real time RT-PCR was also used to quantify changes in the mRNA levels of several genes,such as BMP2,Runt-related transcription factor 2(Runx2),and Osterix(Osx).We also used the Western blot analysis to evaluate the expression of Runx2 and Osx proteins.At last we used the BMP2loxp/loxp mice to isolate the primary calvaria osteoblasts,cultured with Isopsoralen of the best dose for 48 h after the in vitro conditional gene knockout technology,and tested the gene expressions of Runx2 and Osx.And the alkaline Phosphatase(ALP)staining was also performed.Result Isopsoralen(10 μg·mL-1)can promote osteoblast proliferation obviously.From the real time RT-PCR analysis,Isopsoralen can enhance the BMP2 mRNA levels,the effect of 10 μg·mL-1 was the best,and 30 μg·mL-1 followed.In addition,we found that Isopsoralen(10 μg·mL-1)can enhance the Runx2 mRNA levels significantly.We also found that lower doses of Isopsoralen can enhance the Osx mRNA levels,the effect of 30 μg·mL-1 was the best,and 10 μg·mL-1 followed.From the Western blot analysis,low doses of Isopsoralen(10 μg·mL-1 and 30 μg·mL-1)can stimulate the expression of Runx2 protein.Besides,three doses of Isopsoralen can stimulate the expression of Osx protein,and the effect of 10 μg·mL-1 and 30 μg·mL-1 are better.Finally,the results of in vitro conditional gene knockout experiment showed that the overexpression of Runx2 and Osx genes in osteoblasts,as well as ALP staining,induced by Isopsoralen are BMP2 dependent.Conclusions In this study,we firstly demonstrate that Isopsoralen can stimulate osteoblast proliferation and differentiation by mediating BMP2/Runx2/Osx signaling pathway.

Expression and Significance of BMP-7/Smads Signaling Pathway in Hepatic Fibrosis Tissues induced by Common Bile Duct Ligation in Rats / 医学研究杂志

Objective To investigate the expression and significance of BMP7/Smads signaling pathway in hepatic fibrosis tissues induced by common bile duct ligation in rats.Methods Male Wistar rats were simple randomizationally divided into sham operation group,common bile duct ligation group for 7days group,14days group,28days group and 42days group.Common bile duct ligation meth-od was used to establish the cholestatic hepatic fibrosis animal model.hematoxylin-eosin staining and Masson staining were used to ob-serve the hepatic tissues morphology and collagen deposition of the rats in each group.Real-time quantitative polymerase chain reaction(RT-qPCR)technology was used to detect the mRNA expression of hepatic fibrosis indicators,such as α-smooth muscle actin(α-SMA)and fibronectin(FN)in each group.Immunohistochemistry was used to determine the deposition of collagen Ⅰ in the hepatic tis-sues,Western blot was used to detect the proteins expression of FN,collagen Ⅰ,collagen Ⅲ,α-SMA and tissue inhibitor of matrix metalloproteinase(TIMP1)in the hepatic tissues.The expression and localization of BMP-7 and p-Smad1/5/8 proteins in the hepatic tissues were detected by Western blot and Immunohistochemistry.Results Compared with the sham operation group,with the extension of common bile duct ligation time,the degree of hepatic fibrosis gradually increased,the intrahepatic collagen hyperplasia was obvious,and thick fibrous septum were obviously formed in the hepatic issues of BDL groups.The expression of FN,α-SMA,collagen Ⅰ,colla-gen Ⅲ and TIMP1 proteins in the BDL hepatic tissues were significantly increased,the difference were statistically significant(P<0.05).In the hepatic fibrosis tissue,the expression of BMP-7 and p-Smad1/5/8 proteins were significantly increased in the early stage of hepatic fibrosis,while which were decreased with the aggravation of hepatic fibrosis.Compare with the sham operation group,the difference was statistically significant(P<0.05).Conclusion The BMP-7/Smads signaling pathway is significantly activated in the early stage of hepatic fibrosis in rats,which exerted protective effect on bile duct ligation-induced hepatic fibrosis.However,the protec-tive effect of activation of BMP-7/Smads signaling pathway was inhibited with the progression of hepatic fibrotic severity.

Exploration of Mechanism of Xiaonang Tiaojing Decoction in Treating PCOS-IR Model Rats Based on AMH/AMHR ⅡSignaling Pathway / 中国实验方剂学杂志

ObjectiveTo investigate the mechanism of Xiaonang Tiaojing decoction（XNTJD）in improving polycystic ovary syndrome with insulin resistance（PCOS-IR）model rats based on anti-Müllerian hormone（AMH）/AMH type Ⅱ receptor（AMHRⅡ）signaling pathway. MethodForty-eight adult female SD rats were randomly divided into the blank group， model group， XNTJD group（11.4 g·kg-1·d-1） and Diane-35 group（0.21 g·kg-1·d-1）， PCOS-IR model was established by high-fat and high-sugar diet combined with letrozole in rats of all groups except the blank group， rats in the administration groups were given the corresponding dose of drugs by gavage for 15 days with an interval of 1 d every 4 d， normal saline of the same volume was given to the blank group and the model group. Estrous cycle was recorded daily during treatment. At the end of treatment， body weight and Lee's index were recorded， AMH， luteinizing hormone（LH）， LH/follicle stimulating hormone（FSH）， testosterone（T）and estradiol（E2）levels were measured by enzyme-linked immunosorbent assay（ELISA）， fasting plasma glucose（FPG）was measured by glucometer， fasting insulin（FINS） level was measured by radioimmunoassay（RIA）， and the insulin resistance index（HOMA-IR） and insulin sensitivity index（QUICKI）were calculated， triglyceride（TG）and total cholesterol（TC）levels were measured by automatic biochemical analyzer， hematoxylin-eosin（HE）staining was used to observe the morphological changes of the ovary， the levels of AMHRⅡ， bone morphogenetic protein-15（BMP-15）and Smad5 in ovarian tissues were detected by immunohistochemistry（IHC），Western blot was used to analyze the protein expression levels of AMHRⅡ， BMP-15 and Smad5. ResultCompared with the blank group， a large number of leukocytes were observed in the vaginal exfoliated cells of rats in the model group， mainly in diestrus， the levels of body weight， Lee's index， glucose-lipid metabolism indexes（FPG， FINS， HOMA-IR， TG， TC）， AMH and sex hormones（LH， LH/FSH， T）were significantly increased（P<0.01）， and QUICKI and E2 levels were significantly decreased（P<0.01）， there were more cystic bulges on the ovarian surface， more wet weight， more atretic and cystic dilated follicles in the ovarian tissues， and the thickness of granulosa cell layer was reduced without oocytes， the expression level of AMHRⅡ protein in ovarian tissues was significantly increased（P<0.01）， and the expression levels of BMP-15 and Smad5 proteins were significantly decreased（P<0.01）. Compared with the model group， the exfoliated cells in the vagina of rats treated with XNTJD group showed keratinocytes from the 5th to 6th day of treatment， and a stable estrous cycle gradually appeared， body weight， Lee's index， glucose-lipid metabolism indexes（FPG， FINS， HOMA-IR， TG， TC）， AMH and sex hormones（LH， LH/FSH， T）levels were significantly decreased（P<0.05， P<0.01）， QUICKI and E2 levels were significantly increased（P<0.01）， ovarian surface was smoother and lighter in wet weight， oocytes and mature follicles were observed in ovarian tissues， the thickness of granulosa cell layer increased and the morphology was intact， the expression levels of BMP-15 and Smad5 proteins were significantly increased（P<0.01）and the expression level of AMHRⅡ protein was significantly decreased（P<0.01）in ovarian tissues. ConclusionXNTJD may mediate the up-regulation of BMP-15 and Smad5 in ovarian tissues of PCOS-IR rats by down-regulating AMH/AMHRⅡ， thereby improving ovarian function， sex hormones and glucose-lipid metabolism levels in PCOS-IR rats.

Tratamiento farmacológico del estrabismo / Pharmacological treatment of strabismus

La cirugía de la musculatura extraocular ha sido el estándar de atención para tratamiento quirúrgico del estrabismo por más de un siglo. A pesar del gran desarrollo técnico de la cirugía de estrabismo en la actualidad, la utilización de microscopio quirúrgico, el diseño novedoso del instrumental quirúrgico, la calidad de la sutura no reabsorbible; los avances en equipamiento y fármacos anestésicos, la misma no está exenta de complicaciones quirúrgicas, además del tiempo de recuperación que necesita el paciente para reincorporarse a sus actividades sociales, han propiciado una búsqueda permanente del tratamiento farmacológico para el estrabismo. El objetivo de esta revisión bibliográfica es analizar las distintas alternativas farmacológicas disponibles como tratamiento del estrabismo. Para su confección se consultó textos completos y artículos en idiomas español e inglés, disponible en algunas bases de datos. Concluimos que aunque se han estudiado numerosos fármacos, la toxina botulínica que es la más conocida y utilizada mundialmente, seguida de la bupivacaína. Encontramos otros como la IGF I y II (Insuline Growing Factor), capaces de generar un efecto de reforzamiento de la actividad muscular. Y otros que "debilitan" la musculatura extraocular, incluyen la mAb35-Rubicina, BMP4 (Proteína morfogénica ósea). Se continúa su investigación en la actualidad(AU)

Extraocular musculature surgery has been the standard of care for surgical treatment of strabismus for more than a century. Despite the great technical development of strabismus surgery today, the use of a surgical microscope, the novel design of surgical instruments, the quality of the non-absorbable suture; Advances in anesthetic equipment and drugs, it is not exempt from surgical complications, in addition to the recovery time that the patient needs to return to their social activities, have led to a permanent search for pharmacological treatment for strabismus. The objective of this bibliographic review is to analyze the different pharmacological alternatives available as a treatment for strabismus. For its preparation, full texts and articles in Spanish and English languages were consulted, available in some databases. We conclude that although numerous drugs have been studied, botulinum toxin, which is the best known and used worldwide, followed by bupivacaine. We find others such as IGF I and II (Insuline Growing Factor), capable of generating an effect of reinforcing muscle activity. And others that "weaken" MOE include mAb35-Rubicin, BMP4 (Bone Morphogenic Protein). His research is continuing today(AU)

Expression significance of serum miR-27, miR-93, BMP-15 in patients with polycystic ovary syndrome / 中华内分泌外科杂志

Objective:To explore the expression significance of serum MicroRNA-27 (miR-27), MicroRNA-93 (miR-93) and bone morphogenetic protein 15 (BMP-15) in patients with polycystic ovary syndrome (PCOS) .Methods:63 PCOS patients admitted to Yantai Yantaishan Hospital from Aug. 2015 to Jan. 2019 were selected and divided into the obese group (obese PCOS, BMI≥25 kg/m 2, n=22) and the non-obese group (non-obese PCOS, BMI<25 kg/m 2, n=41) according to body mass index (BMI). 30 healthy women with normal physical examination during the same period were selected as the healthy group. Serum miR-27, miR-93 expression levels and bone morphogenetic protein 15 (BMP-15) concentration of the three groups were analyzed, and endocrine metabolism indicators of the three groups were compared. Receiver operating characteristic curve (ROS) was used to evaluate the predictive value of serum miR-27, miR-93 and BMP-15 for PCOS and obese PCOS patients. Results:(1) Endocrine indicators: Compared with the healthy group, FSH was lower in the obese group and the non-obese group, and T, LH/FSH and IR were higher ( P<0.05) ; Compared with the non-obese group, T and IR were higher in the obese group ( P<0.05) ; (2) Serum miR-27, miR-93 and BMP-15: Compared with the healthy group, serum BMP-15 concentration was lower in the obese group and the non-obese group, and serum miR-27, miR-93 expression levels were higher ( P<0.05) ; Compared with the non-obese group, the serum miR-27 and miR-93 expression levels in the obese group were higher ( P<0.05) ; (3) miR-27, miR-93 and BMP-15 had predictive value for PCOS, and the area under the curve of BMP-15 was the highest ( P<0.05) ; (4) miR-27 and miR-93 had predictive value for obese PCOS, and the area under the curve of miR-93 was the highest ( P <0.05) . Conclusions:In addition to significant endocrine index disorders in obese PCOS patients, serum miR-27 and miR-93 are highly expressed and the level of BMP-15 is relatively low. BMP-15 can be used as an effective parameter to assist in the diagnosis of PCOS, and miR-93 can be used as an effective parameter to assist in the diagnosis of obese PCOS.

Influence of photobiomodulation and tricalcium silicate-based cement on viability and dentinogenic activity of cultured human dental pulp stem cells / Influência da fotobiomodulação e cimento de silicato tricálcio na viabilidade e atividade dentinogênica de células tronco cultivadas da polpa de dente humano

Objective: To assess the effect of application of Biodentine (BD), Photobiomodulation (PBM) using 810 nm diode laser and both on the proliferation and odontogenic differentiation of human dental pulp stem cells (HDPSCs). Material and Methods: HDPSCs were collected, isolated, and characterized and then divided into six groups: groups 1, control; groups 2, biodentine (BD); group 3, irradiation at 1 J/cm 2 of 810-nm diode laser; group 4, irradiation at 1 J/cm 2 and culture with BD; group 5, irradiation at 2 J/cm 2, and group 6, irradiation at 2 J/cm 2 and culture with BD. Viability assay was measured through MTT assay and Alkaline phosphatase (ALP) enzyme activity and mRNA levels of RUNX2, collagen 1 (Col-1) and BMP2 were also assessed. Results: Photobiomodulation at 1 and 2 J/cm 2 combined with biodentine significantly promoted HDPSCs proliferation (in MTT assay results) and odontogenic differentiation (through the gene expression of RUNX2, Col-1 and BMP2 levels (p < 0.05). Conclusion: Photobiomodulation at 2 J/cm 2 combined with biodentine enhanced proliferation and odontogenic differentiation of cultured HDPSCs and thus could further be beneficial for dentin regeneration (AU)

Objetivo: Avaliar o efeito da aplicação de Biodentina (BD), Fotobiomodulação (PBM) usando diodo de laser de 810 nm e ambos na proliferação e diferenciação odontogênica de células tronco cultivadas da polpa dental (HDPSCs). Material e Métodos: HDPSCs foram coletadas, isoladas, caracterizadas e então divididas em seis grupos: grupo 1, controle; grupo 2, biodentina (BD); grupo 3, irradiação com diodo de laser a 1 J/cm2 de 810- nm; grupo 4, irradiação a 1 J/cm 2 e cultivo com BD; grupo 5, irradiação a 2 J/cm2, e grupo 6, irradiação a 2 J/cm2 e cultivo com BD. A viabilidade foi mensurada através do teste MTT e a atividade da enzima Fosfatase alcalina (ALP), e níveis de RNAm de RUNX2, de colágeno 1 (Col-1) e de BMP2 foram também mensurados. Resultados: Fotobiomodulação a 1 e 2 J/cm 2 combinada com biodentina promoveu significativa proliferação de HDPSCs (nos resultados do teste MTT) e diferenciação odontogênica (através da expressão genética dos níveis de RUNX2, Col-1 e BMP2 (p < 0.05)). Conclusão: Fotobiomodulação a 2 J/cm2 combinada com biodentina aumentou a proliferação e diferenciação odontogênica de HDPSCs cultivadas e dessa forma poderia ser benéfica para a regeneração dentinária. (AU)

The long-term effect of hip replacement and vitamin D in the treatment of elderly osteoporotic femoral neck fractures and its effect on the level of BMP-7/25- (OH) -D-3 / 中华内分泌外科杂志

Objective:To explore the long-term effect of hip replacement and vitamin D in treatment of elderly osteoporotic femoral neck fractures and the transfection of bone morphogenetic protein-7 (BMP-7) /25 hydroxy vitamin D3[ (25- (OH) ) -D3] level.Method:Data of 108 elderly osteoporotic femoral neck fracture patients admitted from Jan. 2018 to Jan. 2020 were selected, and they were divided them into the observation group (hip replacement adjuvant vitamin D treatment) and control group (hip replacement treatment) , 54 cases in each group. All subjects were followed up for 1 year to observe the long-term treatment efficacy and SF-36 scores of the two groups of patients. Before treatment and 1, 3, 6, and 12 months after treatment, the differences in the changes in lumbar and hip bone mineral density, hip joint Harris score, BMP-7, and 25- (OH) -D3 levels were compared between the two groups.Results:12 months after treatment, the long-term treatment effect of the observation group was significantly higher than that of the control group ( P<0.05) ; 1, 3, 6, 12 months after treatment, for the lumbar and hip bone mineral density, SF-36 score, hip Harris score, BMP-7, 25- (OH) -D3 levels of the two groups of patients over time, the degree of increase was not equal, but the treatment group had the greater degree of improvement ( P<0.01) . Conclusion:Hip joint replacement and vitamin D have a good long-term effect in treatment of elderly osteoporotic femoral neck fractures, which can significantly increase bone density and improve BMP-7 and 25- (OH) -D3 levels.

Expression and significance of TGF-β1, p38MAPK and BMP-7 protein in liver specimens of patients with alveolar hepatic echinococcosis / 中国血吸虫病防治杂志

Objective To detect the expression of transforming growth factor-β1 (TGF-β1), p38MAPK and bone morphogenetic protein-7 (BMP-7) protein in the liver specimens of patients with hepatic alveolar echinococcosis, and to investigate the potential role of TGF-β1, p38MAPK and BMP-7 protein in hepatic fibrosis caused by hepatic alveolar echinococcosis. Methods A total of 20 patients with hepatic alveolar echinococcosis were enrolled as study subjects, and hepatic specimens were sampled from the sites within 0.5 cm (Group A) and 0.5 to 1.5 cm from hepatic alveolar echinococcosis lesions (Group B), while normal liver specimens sampled from the sites 2 cm and greater from hepatic alveolar echinococcosis lesions served as controls (Group C). The fibrosis of liver specimens was pathological examined using HE and Masson staining, and the expression of TGF-β1, p38MAPK and BMP-7 protein was quantified in liver tissues using Western blotting. The associations of TGF-β1, p38MAPK and BMP-7 protein expression with hepatic fibrosis were assessed. Results HE staining showed the malaligned structure of hepatocytes and destruction of the structure of hepatic lobules at various degrees in liver specimens in groups A and B, with hepatocyte degeneration, atrophy and necrosis, hyperplasia of fibrous tissues and eosinophilic granulocyte infiltration seen, while no abnormal pathological alterations of liver tissues, normal hepatocyte structure and morphology and uniform size, no malaligned structure of hepatocytes, clear structure of hepatic lobules, no or mild hepatocyte degeneration or necrosis, and no eosinophilic granulocyte infiltration were seen in Group C. Masson staining showed that there was hyperplasia of multiple fibrous connective tissues in the liver portal areas in groups A and B, with fibrosis seen in hepatic lobules, while no obvious pathological changes were seen in Group C. There were significant differences seen in TGF-β1 (P < 0.001), p38MAPK (P < 0.01) and BMP-7 protein (P < 0.05) expression in liver tissues in groups A, B and C, and higher TGF-β1, p38MAPK and BMP-7 protein expression was quantified in groups A and B than in Group C (all P values < 0.05), while greater TGF-β1, p38MAPK and BMP-7 protein expression was detected in Group B than in Group C (all P values < 0.05). The expression of TGF-β1, p38MAPK and BMP-7 protein correlated positively with the severity of hepatic fibrosis (r = 0.866, 0.702 and 0.801, all P values < 0.05), and there were significant differences in TGF-β1 (F = 72.580, P < 0.01), p38MAPK (χ2 = 31.705, P < 0.01) and BMP-7 protein expression (χ2 = 48.388, P < 0.01) among liver tissues with different degrees of fibrosis. The TGF-β1 protein expression correlated positively with p38MAPK and BMP-7 protein expression (r = 0.607 and 0.702, both P values < 0.001), and the BMP-7 protein expression also correlated positively with p38MAPK protein expression (r = 0.456, P < 0.001). Conclusion The interaction among TGF-β1, p38MAPK and BMP-7 jointly participates in the development of hepatic fibrosis induced hepatic alveolar echinococcosis.