Objective To screen the active components of Bushen Huoxue Decoction ( BSHXD) involved in promoting the proliferation of bone marrow mesenchymal stem cells ( MSCs). Methods BSHXD and its subdivisions were extracted with petroleum ether, ethyl acetate, water-free ethanol and water respectively. MSCs were isolated and cultured by the bone marrow adherent method. At the third passage, MSCs were identified by the specific surface markers with immunofluorescence, and their osteogenic and adipogenic differentiation were tested by alizarin red staining and oil red “O” staining. After treated with the extracts of BSHXD and its subdivisions at gradient concentrations for 24 hours, cell viability was detected by methyl thiazolyl tetrazolium (MTT) assay for the screening of active components and optimal concentration. MTT assay was used to describe the growth curve of MSCs treated with the most effective components, and cell cycle was analyzed by flow cytometry. Results Compared with the blank control group, the extracts of BSHXD and its subdivisions could protect MSCs from death to various degrees. Of all the extracts, the ethyl acetate extract of Bushen Division ( BSD) , ethyl acetate extract of BSHXD, ethyl acetate extract of Huoxue Division ( HXD) had the strongest effect, and the effect was dose-dependent, 100 μg/mL being the optimal active concentration while having no any cytotoxic reaction. The results of MTT assay revealed that BSD extracts promoted the proliferation of MSCs significantly and was the most effective component, and then came BSHXD. The results of flow cytometry indicated that BSD extract had the most strongest effect on increasing the amount of MSCs at proliferative phase, and then came BSHXD. Conclusion BSD ethyl acetate extract is the active component of BSHXD for promoting the proliferation of MSCs, showing an effect on increasing the proportion of MSCs at proliferative phase.

BACKGROUND:Mechanical stress plays an important role in the progressive collapse in osteonecrosis of the femoral head.Therefore,the bone structure of weight-bearing area may be the risk factor for collapse.OBJECTIVE:To investigate the relationship between the bone preservation of weight-bearing area and collapse progression in necrotic femoral head.METHODS:Eighty-seven patients (102 hips) with ARCO stage Ⅱ osteonecrosis of the femoral head,diagnosed by MRI,were enrolled.All patients underwent a natural progression.Collapse and bone involvement of the weight-bearing area were viewed by anteroposterior and frog-leg lateral radiographs.According to the location of the necrotic lesion on the anterolateral portion of the femoral head,the necrosis was divided into three types:type 1,the posteromedial and central portions;type 2,part of the anterolateral portion;type 3,the entire anterolateral portion.The collapse rate and the time to collapse in different types were assessed.RESULTS AND CONCLUSION:(1) All patients were followed up for 3-58 months.(2) Of the 60 hips with collapse,46 (76.7％) hip collapse was identified on anteroposterior radiograph.On frog-leg lateral radiograph showing collapsed femoral head could be identified in 57 (95.0％) hips,which was significantly different (P ＜ 0.01).(3) In all 102 hips,the collapse rate in type 3 osteonecrosis of the femoral head was significantly higher than that of type 2 osteonecrosis of the femoral head (P ＜ 0.001),and the time to collapse was markedly shortened.None collapse occurred in all six hips with type Ⅰ osteonecrosis of the femoral head during follow-up.(4) In summary,preservation of anterolateral portion is associated with potential collapse progression in necrotic femoral head.

Objective To establish a PCR-based capillary electrophoresis(PCR/CE)to detect Survival Motor Neuron 1(SMN1)and Survival Motor Neuron 2(SMN2)genes and to evaluate its performance.Methods PCR/CE and Multiplex Ligation-dependent Probe Amplification(MLPA)for SMA gene diagnosis were used to blindly test the samples in sync.The performance of PCR/CE was assessed using MLPA results as the standard.Results A total of 336 samples were included in this study,consisting of 50 homozygous deletion types(14.9%),65 heterozygous deletion types(19.3%),and 221 non-deletion types(65.8%).The results of PCR/CE for detect-ing SMN1 and SMN2 copy numbers(0,1,2,3,≥4)were in complete agreement with the results of the MLPA.Conclusions PCR/CE for gene testing related to SMA could accurately detect copy numbers of exon 7 and exon 8 of the SMN1 and SMN2 genes(0,1,2,3,≥4).

BACKGROUND:Compound Shengmai Chenggu capsule has good therapeutic effects on early steroid-induced osteonecrosis of the femoral head,but the exact mechanism of treatment is not fully understood. OBJECTIVE:To observe the effect of compound Shengmai Chenggu capsule on fucosyltransferase 8,osteogenic gene and Wnt/β-catenin in bone tissue of rats with steroid-induced osteonecrosis of the femoral head. METHODS:Sixty Sprague-Dawley rats were randomized into blank group,model group,low-,middle-,and high-dose drug groups(n=12 per group).In the latter four groups,animal models of steroid-induced osteonecrosis of the femoral head were established by subcutaneous injection of imiquimod(once every 2 weeks,2 times in total)and gluteal muscle injection of methylprednisolone(once a week,4 times in total).The low-,middle-and high-dose drug groups were given 1.89,3.78 and 7.56 g/kg per day compound Shengmai Chenggu capsule solution by gavage respectively on the second day after the last modeling.The same amount of saline was given by gavage to the model group.Administration lasted 8 weeks.After the administration,micro-CT scan,histological staining,compression test,RT-qPCR and western blot were performed on the femoral head. RESULTS AND CONCLUSION:Micro-CT scan results showed that compared with the blank group,trabecular volume fraction,trabecular number and trabecular thickness were significantly decreased(P<0.05),while trabecular separation was increased in the model group(P<0.05).Compared with the model group,the compound Shengmai Chenggu capsule could increase trabecular volume fraction,trabecular number and trabecular thickness(P<0.05),and decrease trabecular separation(P<0.05)in a dose-dependent manner.Hematoxylin-eosin staining results showed that compared with the model group,the rate of empty bone lacunae was reduced in a dose-dependent group in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups(P<0.05).Immunohistochemical staining results showed that compared with the blank group,the protein expression of fucosyltransferase 8,Runx2 and bone morphogenetic protein 2 was reduced in the model group(P<0.05);compared with the model group,the compound Shengmai Chenggu capsule increased the protein expression of fucosyltransferase 8,Runx2 and bone morphogenetic protein 2 in a dose-dependent manner(P<0.05).Results from the compression test showed that there was a dose-dependent increase in the maximum load and elastic modulus of the femoral head in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups compared with the model group(P<0.05).RT-qPCR and western blot results showed that the mRNA and protein expressions of fucosyltransferase 8,Runx2,alkaline phosphatase,osteocalcin,osteoblast-specific transcription factor and bone morphogenetic protein 2 were decreased in the model group compared with the blank group(P<0.05);compared with the model group,there was a dose-dependent increase in the mRNA and protein expressions of the above indicators in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups compared with the model group(P<0.05).Compared with the blank group,the mRNA and protein expression of Wnt2,low-density lipoprotein receptor-related protein 5 and β-catenin were decreased(P<0.05)and the mRNA and protein expressions of glycogen synthase kinase 3β were increased(P<0.05)in the model group;compared with the model group,there was a dose-dependent increase in the mRNA and protein expressions of Wnt2,low-density lipoprotein receptor-related protein 5 and β-catenin(P<0.05)but a dose-dependent decrease in the mRNA and protein expressions of lycogen synthase kinase 3β(P<0.05)in the low-,middle-,and high-dose compound Shengmai Chenggu capsule groups.To conclude,the mechanism by which the compound Shengmai Chenggu capsule treats steroid-induced osteonecrosis of the femoral head may activate the Wnt/β-catenin signaling pathway through the up-regulation of fucosyltransferase 8,thereby promoting bone formation.

BACKGROUND:The majority of studies on developmental dysplasia of the hip focus on hip malformations,but there are few reports on the effects of acetabular dysplasia on the spine. OBJECTIVE:To investigate the compensation of spinopelvic parameters in coronal and sagittal views in patients with developmental dysplasia of the hip,and to explore the correlation between acetabular development and spinopelvic parameters. METHODS:A total of 101 patients with developmental dysplasia of the hip admitted to the Third Affiliated Hospital of Guangzhou University of Chinese Medicine from January 2018 to June 2022 were selected as the trial group,and 114 healthy subjects were selected as the control group during the same period.The spinopelvic parameters of the subjects were measured through the full-length X-ray films of the coronal and sagittal spines:lumbar lordosis,anterior pelvic tilt,thoracolumbar kyphosis,Cobb angle,and the distance between the C7 plumb line and the center sacral vertical line,sacral slope,pelvic incidence,and thoracic kyphosis.The differences in spinopelvic parameters were compared between the two groups.In addition,the differences in spinopelvic parameters in patients with unilateral,bilateral and different Crowe classifications of developmental dysplasia of the hip were compared.Pearson correlation analysis was used to explore the correlation between Sharp angle and spinopelvic parameters. RESULTS AND CONCLUSION:(1)In the sagittal view,the lumbar lordosis in the trial group was significantly lower than that in the control group(P<0.05).The pelvic tilt and kyphosis angle of the thoracolumbar segment in the trial group were significantly greater than those in the control group(P<0.05).In the coronary position,the Cobb angle and the distance between the C7 plumb line and center sacral vertical line in the trial group were significantly greater than those in the control group(P<0.05).There was no significant difference in the remaining spinopelvic parameters between the two groups(P>0.05).(2)The lumbar lordosis of patients with bilateral developmental dysplasia of the hip was significantly lower than that of patients with unilateral developmental dysplasia of the hip(P<0.05).The pelvic tilt,thoracolumbar kyphosis,Cobb angle and the distance between the C7 plumb line and center sacral vertical line in bilateral developmental dysplasia of the hip patients were significantly greater than those in unilateral developmental dysplasia of the hip patients(P<0.05).(3)The lumbar lordosis decreased with the increase of Crowe classification severity(P<0.05).The pelvic tilt increased with the severity of the Crowe classification(P<0.05).(4)Pearson correlation analysis showed that Sharp angle was negatively correlated with lumbar lordosis(P<0.05),while Sharp angle was positively correlated with anterior pelvic tilt,Cobb angle,C7 plumb line and center sacral vertical line(P<0.05).(5)It is concluded that the pelvic tilt,thoracolumbar kyphosis,Cobb angle and the distance between the C7 plumb line and center sacral vertical line increase,while lumbar lordosis decreases in developmental dysplasia of the hip patients.The degree of acetabular dysplasia was significantly correlated with lumbar lordosis,pelvic tilt,Cobb angle,C7 plumb line and center sacral vertical line.