ObjectiveTo unveil the mechanism of Jianpi Huogu prescription （JPHGP） in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head （SONFH） by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal， model， low-， medium-， and high-dose （2.5， 5， 10 g·kg^-1， respectively） JPHGP， and Jiangushengwan （1.53 g·kg^-1） groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg^-1 on days 1 and 2， and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg^-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection， and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head， and the number of adipocytes， the rate of empty bone lacunae， and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein （LDL）， high-density lipoprotein （HDL）， apolipoprotein A1 （ApoA1）， and apolipoprotein B （ApoB）. At the same time， the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed， and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking， immunohistochemistry， and Western blot. ResultsCompared with the normal group， the model group showcased thinning of the femoral head， trabecular fracture， karyopyknosis， subchondral cystic degeneration， increases in the number of adipocytes and the rate of empty bone lacunae （P<0.01）， a reduction in the trabecular area （P<0.01）， decreases in BMD， Tb.Th， Tb.N， and BV/TV， and increases in Tb.Sp and BS/BV （P<0.01）. Compared with the model group， the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae， an increase in the trabecular area （P<0.05， P<0.01）， rises in BMD， Tb.Th， Tb.N， and BV/TV， and decreases in Tb.Sp and BS/BV （P<0.05， P<0.01）. Compared with the normal group， the model group showcased raised serum levels of TG， TC， LDL， and ApoB and lowered serum levels of HDL and ApoA1 （P<0.01）. Compared with the model group， the JPHGP groups had lowered serum levels of TG， TC， LDL， and ApoB （P<0.05， P<0.01） and a risen serum level of ApoA1 （P<0.05， P<0.01）. Moreover， the serum level of HDL in the high-dose JPHGP group increased （P<0.01）. A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head， and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation， lipids， apoptosis， and osteoclast differentiation. Molecular docking， immunohistochemistry， and Western blot results showed that compared with the normal group， the model group displayed increased content of 5-lipoxygenase （5-LO） and peroxisome proliferator-activated receptor γ （PPARγ） in the femoral head （P<0.01）. Compared with the model group， medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ （P<0.05， P<0.01）. ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.

ObjectiveTo unveil the mechanism of Jianpi Huogu prescription （JPHGP） in ameliorating the dyslipidemia of steroid-induced osteonecrosis of the femur head （SONFH） by oxylipidomics combined with transcriptomics. MethodsSixty SD rats were assigned into normal， model， low-， medium-， and high-dose （2.5， 5， 10 g·kg^-1， respectively） JPHGP， and Jiangushengwan （1.53 g·kg^-1） groups. Lipopolysaccharide was injected into the tail vein at a dose of 20 μg·kg^-1 on days 1 and 2， and methylprednisolone sodium succinate was injected at a dose of 40 mg·kg^-1 into the buttock muscle on days 3 to 5. The normal group received an equal volume of normal saline. Drug administration by gavage began 4 weeks after the last injection， and samples were taken after administration for 8 weeks. Hematoxylin-eosin staining was conducted to reveal the histopathological changes of the femoral head， and the number of adipocytes， the rate of empty bone lacunae， and the trabecular area were calculated. Micro-computed tomography was used for revealing the histological and histomorphometrical changes of the femoral head. Enzyme-linked immunosorbent assay was employed to measure the serum levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein （LDL）， high-density lipoprotein （HDL）， apolipoprotein A1 （ApoA1）， and apolipoprotein B （ApoB）. At the same time， the femoral head was collected for oxylipidomic and transcriptomic detection. The differential metabolites and differential genes were enriched and analyzed， and the target genes regulating lipid metabolism were predicted. The predicted target proteins were further verified by molecular docking， immunohistochemistry， and Western blot. ResultsCompared with the normal group， the model group showcased thinning of the femoral head， trabecular fracture， karyopyknosis， subchondral cystic degeneration， increases in the number of adipocytes and the rate of empty bone lacunae （P<0.01）， a reduction in the trabecular area （P<0.01）， decreases in BMD， Tb.Th， Tb.N， and BV/TV， and increases in Tb.Sp and BS/BV （P<0.01）. Compared with the model group， the JPHGP groups showed no obvious thinning of the femoral head or subchondroidal cystic degeneration. The high- and medium-dose JPHGP groups presented declines in the number of adipocytes and the rate of empty bone lacunae， an increase in the trabecular area （P<0.05， P<0.01）， rises in BMD， Tb.Th， Tb.N， and BV/TV， and decreases in Tb.Sp and BS/BV （P<0.05， P<0.01）. Compared with the normal group， the model group showcased raised serum levels of TG， TC， LDL， and ApoB and lowered serum levels of HDL and ApoA1 （P<0.01）. Compared with the model group， the JPHGP groups had lowered serum levels of TG， TC， LDL， and ApoB （P<0.05， P<0.01） and a risen serum level of ApoA1 （P<0.05， P<0.01）. Moreover， the serum level of HDL in the high-dose JPHGP group increased （P<0.01）. A total of 19 different metabolites of disease set and drug set were screened out by oxylipidomics of the femoral head， and 119 core genes with restored expression were detected by transcriptomics. The enriched pathways were mainly concentrated in inflammation， lipids， apoptosis， and osteoclast differentiation. Molecular docking， immunohistochemistry， and Western blot results showed that compared with the normal group， the model group displayed increased content of 5-lipoxygenase （5-LO） and peroxisome proliferator-activated receptor γ （PPARγ） in the femoral head （P<0.01）. Compared with the model group， medium- and high-dose JPHGP reduced the content of 5-LO and PPARγ （P<0.05， P<0.01）. ConclusionJPHGP can restore the levels of oxidized lipid metabolites by regulating the 5-LO-PPARγ axis to treat SONFH in rats. Relevant studies provide experimental evidence for the efficacy mechanism of JPHGP in the treatment of SONFH.

OBJECTIVE To investigate the effects of different sterilization methods on 3D-printed polylactic acid(PLA)guide plates for orthopedic surgery,and to analyze their physical properties,microbial contamination,structural performance and bio compatibility after sterilization.METHODS PLA guide plates for orthopedic surgery were prepared with 3D printing technology and divided into a blank group,a hydrogen peroxide plasma steriliza-tion group,an ethylene oxide sterilization group and a pressure steam sterilization group.Before and after sterili-zation,the changes in volume and weight of the surgical guide plates were measured,microbial detection was con-ducted,structural changes were observed with a laser scanning confocal microscope,and cell co-culture was con-ducted to evaluate biocompatibility,thereby the effects of different sterilization methods were assessed.RESULTS The three sterilization methods had no significant effect on the volume and weight of PLA surgical guide plates.Microbial detection showed that all three sterilization methods were effective in killing bacteria,and bacte-rial cultures were negative.Laser confocal scanning microscopy revealed that sterilization treatment caused certain changes to the microstructure of the surgical guide plates,but high-temperature sterilization had a more pro-nounced effect on the deformation of the guide plate edges.Cell co-culture results indicated that the surgical guide plates treated with the three sterilization methods exhibited acceptable cytotoxicity and had little effect on cell pro-liferation.CONCLUSIONS All three sterilization methods cause microstructural changes to the surgical guide plate.Among them,pressure steam sterilization significantly deforms the structure of the guide plate,directly af-fecting its precise positioning of mutual spatial distances,angular relationships and orientation during surgery.Al-though hydrogen peroxide plasma sterilization does not cause significant deformation,ethylene oxide sterilization has the least impact on material properties and structural stability while ensuring the sterilization effect of PLA surgical guide plates.

OBJECTIVE To investigate the effects of different sterilization methods on 3D-printed polylactic acid(PLA)guide plates for orthopedic surgery,and to analyze their physical properties,microbial contamination,structural performance and bio compatibility after sterilization.METHODS PLA guide plates for orthopedic surgery were prepared with 3D printing technology and divided into a blank group,a hydrogen peroxide plasma steriliza-tion group,an ethylene oxide sterilization group and a pressure steam sterilization group.Before and after sterili-zation,the changes in volume and weight of the surgical guide plates were measured,microbial detection was con-ducted,structural changes were observed with a laser scanning confocal microscope,and cell co-culture was con-ducted to evaluate biocompatibility,thereby the effects of different sterilization methods were assessed.RESULTS The three sterilization methods had no significant effect on the volume and weight of PLA surgical guide plates.Microbial detection showed that all three sterilization methods were effective in killing bacteria,and bacte-rial cultures were negative.Laser confocal scanning microscopy revealed that sterilization treatment caused certain changes to the microstructure of the surgical guide plates,but high-temperature sterilization had a more pro-nounced effect on the deformation of the guide plate edges.Cell co-culture results indicated that the surgical guide plates treated with the three sterilization methods exhibited acceptable cytotoxicity and had little effect on cell pro-liferation.CONCLUSIONS All three sterilization methods cause microstructural changes to the surgical guide plate.Among them,pressure steam sterilization significantly deforms the structure of the guide plate,directly af-fecting its precise positioning of mutual spatial distances,angular relationships and orientation during surgery.Al-though hydrogen peroxide plasma sterilization does not cause significant deformation,ethylene oxide sterilization has the least impact on material properties and structural stability while ensuring the sterilization effect of PLA surgical guide plates.

ObjectiveFrom the perspective of energy metabolism， the mechanism of Osteoking （OK） in the treatment of myofascial pain syndrome （MPS） was revealed through systems biology prediction combined with holistic animal experimental validation methods. MethodFirstly， the key targets of MPS and their related molecular mechanisms were predicted by the systems biology method， and the core network targets were screened. Then， the network-predicted targets were verified by animal experiments. Specifically， 60 SD rats were randomly divided into normal group， model group， low， medium， and high dose OK groups （0.66， 1.31， 2.63 mL·kg^-1）， and positive celecoxib group （21 mg·kg^-1）. The MPS model was established by beating combined with a centrifugal exercise method for eight weeks. Except for two days after modeling， the intervention of OK or celecoxib was performed. After the completion of the model， the drug was administered for two weeks. The histopathological changes of trigger point muscle tissue were observed by hematoxylin-eosin staining. The content/activity of Na-K-ATP enzyme （Na⁺-K⁺-ATPase）， Ca²⁺ pump （Ca²⁺ATPase）， Ca²⁺， lactate dehydrogenase （LDH）， glutathione （GSH）， malondialal （MDA）， superoxide dismutase （SOD）， cyclic adenosine phosphate （cAMP）， and protein kinase A （PKA） in serum and/or trigger point muscle tissue in MPS rats was detected by enzyme-linked immunosorbent assay. Protein expression levels of PKA and the peroxisome proliferator-activated receptor γ coactivator 1α （PGC1α） in MPS rats were detected by immunohistochemistry. The protein expression levels of PKA， PGC1α， and mitochondrial transcription factor A （TFAM） in MPS rats were detected by Western blot. ResultThe network prediction results suggest that OK acts on the key target of energy metabolism related to the occurrence and development of MPS and may participate in the activation of the cAMP/PKA/PGC1α signaling pathway. The experimental validation results show that compared with the normal group， contracture nodules and disordered arrangement of muscle fibers appear in the trigger point muscle tissue of MPS rats. Na⁺-K⁺-ATPase， Ca²⁺ATPase， SOD activity， Ca²⁺， and GSH contents in serum and/or trigger point muscle tissue are significantly decreased （P<0.01）. Both LDH activity and MDA contents are significantly increased （P<0.01）， and the protein expression levels of cAMP， PKA， PGC1α， and TFAM are significantly decreased （P<0.01）. Compared with the model group， OK improves the histopathological morphology of trigger point muscle fibers in MPS rats， and after the intervention of OK， Na⁺-K⁺-ATPase， Ca²⁺ATPase， SOD activity， Ca²⁺， and GSH contents in serum and/or trigger point muscle tissue in MPS rats are significantly increased （P<0.05， P<0.01）. LDH activity and MDA contents are significantly reduced （P<0.05， P<0.01）. The protein expression levels of cAMP， PKA， PGC1α， and TFAM are significantly increased （P<0.05， P<0.01）. ConclusionThe mechanism of OK's intervention in MPS rats may be related to its effective activation of the cAMP/PKA/PGC1α signaling pathway， thus promoting mitochondrial energy metabolism and trigger point muscle fiber damage repair in muscle cells.

Objective:To investigate the correlation between sleep duration, sleep timing and the risk of osteoporosis in postmenopausal women, to identify contributing mechanisms and guide the prevention and treatment of osteoporosis.Methods:A total of 5 449 postmenopausal women were included in this study. All participants completed questionnaires, medical examinations, blood test and the measurement of bone mineral density using calcaneal quantitative ultrasonography. After adjusting for potential confounders, logistic regression model was used to assess the association of sleep duration, sleep timing with the risk of osteoporosis. Results:In postmenopausal women, there were significant differences in sleep duration and timing among groups with different risk of osteoporosis( P<0.05). After controlling ages, BMI, diabetes, hypertension, dyslipidemia, smoking, alcohol consumption and physical activity, sleep duration was correlated with the risk of osteoporosis, long sleep duration(≥9 h)increased the risk of osteoporosis( OR=1.39, 95% CI 1.17-1.65, P<0.05)compared with the group with sleep duration of 7~8 hours. In analysis of the combined effect of sleep duration and sleep time on the risk of osteoporosis, compared with normal sleep duration(7-8 h)and normal sleep timing(22: 00-23: 00), long sleep duration(≥9 h)and normal sleep timing(22: 00-23: 00)increased the risk of osteoporosis( OR=1.38, 95% CI 1.01-1.87, P<0.05), which was higher in the group of long sleep duration(≥9 h)and late sleep timing(≥23: 00; OR=1.43, 95% CI 1.01-2.01, P<0.05). Conclusion:Long sleep duration(≥9 h)and late sleep timing(≥22: 00)are risk factors for the increased risk of osteoporosis in postmenopausal women, the late sleep timing leads to the higher risk.

Objective To investigate the association between genetic polymorphisms of Dectin-2 and pulmonary cryptococcosis.Methods A total of 134 non-human immunodeficiency virus (HIV)patients with pulmonary cryptococcosis and 464 healthy controls were included in this case control study.The peripheral leucocyte DNA was extracted and genotyping was performed by multiplex SNaPshot technology.The single nucleotide polymorphism (SNP)of rs11045418 located at 5′-flanking locus of Dectin-2 gene was genotyped.Patients without predisposing conditions were compared independently.The differences of gene polymorphism distributions compared between pulmonary patients and healthy control, and between patients without predisposing conditions and healthy control.All data were analyzed withχ2 tests.Results Among the total 134 patients,82 patients had no predisposing factors.Thirty two patients met the proven diagnosis criteria and 102 patients were probable pulmonary cryptococcosis.According to the site of infection, 72 patients had local infection in lungs and 62 patients had disseminated cryptococcosis.Three samples failed in genotyping,one of which was a patient without predisposing factor.Compared with the control group,there was a trend of increasing proportion of heterozygote rs11045418 CT in the 131 pulmonary cryptococcosis patients (59% vs 50%,P =0.069,OR=1.44,95%CI :0.97-2.13),and the heterozygote was significantly increased in 81 patients without predisposing conditions(64% vs 50%,P =0.017,OR= 1 .82,95 %CI :1 .11 -2.95 ).No significant difference of genotype distribution was found between the local and disseminated infection patients.Conclusion Our study shows that rs11045418 CT heterozygote in Dectin-2 is associated with the susceptibility of pulmonary cyrptococcosis among non-HIV-infected Chinese patients,which indicated that the change of Dectin-2 receptor may play a role in the pathogenesis of pulmonary cyrptococcosis.

Objective To investigate the relationship between the expression of β-catenin and drug-resistance mechanism of choriocarcinoma according to the expression of β-catenin in JEG-3 cells (human choriocarcinoma cell line) and drug resistant JEG-3/VP16 cells.Methods The mRNA and protein expressions of β-catenin were analyzed with reverse transcription-polymerase chain reaction(RT-PCR) and Western blotting.Flow cytometry was used to determine the percentages of β-catenin-positive cells in the two choriocarcinoma cell lines.Results Both drug resistant choriocarcinoma cells and drag sensitive cells were found to express β-catenin; but the expression of β-catenin mRNA (1.43 ±0.24) and protein(1.49 ±0.17)in drug resistant choriocarcinoma cells was found much higher than that in drug sensitive cells(0.65 ±0.14,0.66 ±0.16,P ＜0.01).And according to detect by flow cytometry,we found the number of β-catenin-positive cells in JEG-3/VP16 cells [(40.13 ±5.17) ％] was much more than that in JEG-3 cells [(13.15 ± 1.48) ％,P ＜ 0.01].Conclusions β-catenin was highly expressed in the drug resistant choriocarcinoma cell line (JEG-3/VP16).It indicates β-catenin might be involved in the drug resistance mechanism of choriocareinoma.

Objective To explore the relationships among attachment,parental attachment and parental rearing style of adolescent depression.Methods 36 outpatients and hospital patients in Anhui Mental Health Center with depression and their parents were involved as the case group,33 age-,gender-,and education-matched high school students and their parents as the normal controls.The case group and control group were assessed with Experiences in Close Relationships Inventory (ECR) and Parental Rearing Pattern Questionnaire (EMBU).Results ① The score of attachment avoidance and anxiety dimension in the depression group were obviously higher than that of the control group(73.19 ± 16.63 vs 66.25 ± 12.91,62.76 ± 13.90 vs 51.97 ± 13.69,P ＜ 0.01) ;the score of mother's attachment anxiety in the depression group were significantly higher than that of the control group (72.06 ± 14.23 vs 57.42 ± 12.81),and there were significant differences between two groups (P ＜ 0.01).②The attachment avoidance was positively correlated with parental attachment avoidance in the depression group (r =0.534,r=0.488 ; P＜ 0.01).The attachment anxiety dimension was positively correlated with mother's attachment anxiety in depression group (r =0.532,P ＜ 0.01).Attachment avoidance was negatively correlated with parental warmness(r =-0.406,r =-0.462,P ＜ 0.01),however attachment avoidance had positive correlation with father's punishment and mother's rejection in depression group (r=0.395,r=0.468; P＜0.05).Father's attachment avoidance was negatively correlated with father's warmness(r =-0.527,P＜ 0.01),and mother's attachment avoidance was negatively correlated with mother's warmness (r =-0.491,P ＜ 0.01),which was positively correlated with mother's rejection (r =0.392,P ＜ 0.05).③ Regression analysis showed that mother's warmness rearing style had full mediating effect on the relationship between mother's attachment avoidance and attachment avoidance in depression group,and then mother's refused rearing style partly mediated the relationship between them.But father's warmness didn(t) have mediating effect on the relationship between father's attachment avoidance and attachment avoidance in depression group.Conclusion The attachment pattern is insecure in the depression group.Parenting attachment and rearing style significantly influence on adolescent depression attachment patterns,and mother's rearing style has mediating effect on the relationship between mother's attachment and depression attachment.