OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.

OBJECTIVE To study and analyse the main components of Tongluo Muzu Powder transdermal absorption solution,as well as to investigate its mechanism of action in the treatment of osteoarthritis of the knee.METHODS The transdermal absorption solution of Tongluo Muzu Powder was extracted by Franz vertical diffusion cell,and its components were analyzed by UHPLC-MS/MS.Then TCMSP,Swiss target prediction,Gene Cards and other databases were introduced to predict the possible targets of the effec-tive components of transdermal absorption solution for the prevention and treatment of knee osteoarthritis,and Cytoscape software was used to construct the"drug-component-disease-target"visual network,and then the protein interaction network was obtained through the string database to find the core targets.Finally,AutoDock and PyMOL were used to verify the molecular docking of main active in-gredients and targets,and the gene ontology(GO)function analysis of core genes and the enrichment analysis of Kyoto Encyclopedia of genes and genomes(KEGG)pathway were conducted.Primary mouse chondrocytes were extracted,and the predicted targets of net-work pharmacology were verified by ELISA,Western blot,qPCR,etc.RESULTS A total of 48 effective components of the transder-mal absorption solution of Tongluo Muzu Powder and 88 possible targets for the treatment of knee osteoarthritis were screened out.The results of enrichment analysis showed that it was mainly involved in the process of apoptosis and oxidative stress.Molecular docking suggested that the main active ingredients and targets had good binding ability.The results showed that the lyophilized powder of the transdermal absorption solution of Tongluo Muzu Powder could reduce the concentration of inflammatory factors in the supernatant of chondrocytes after LPS intervention in a dose-dependent manner(P<0.05,P<0.01).The protein expression of p-PI3K/PI3K,p-AKT/AKT,MMP13 and p53 was decreased,and the protein expression of collagen Ⅱ was increased(P<0.05,P<0.01).At the same time,the mRNA expression of MMP13 and p53 was decreased and the mRNA expression of collagen Ⅱ was increased(P<0.05,P<0.01).In addition,it reduced the fluorescence intensity of TUNEL staining in cells(P<0.01).CONCLUSION By combining UH-PLC-MS/MS technology with network pharmacology,the main active ingredients of Tongluo Muzu Powder transdermal absorption solu-tion are preliminarily understood,and its potential mechanism of action in the treatment of knee osteoarthritis are predicted.

BACKGROUND: Gestational diabetes mellitus (GDM) brings health issues for both mothers and offspring, and GDM prevention is as important as GDM management. It was shown that a history of GDM was significantly associated with a higher maternal risk for GDM recurrence. The incidence of GDM recurrence was unclear because of the incidence of second-child was low before 2016 in China. We aim to investigate the prevalence of GDM recurrence and its associated high-risk factors which may be useful for the prediction of GDM recurrence in China. METHODS: A retrospective study was conducted which enrolled participants who underwent regular prenatal examination and delivered twice in the same hospital of 18 research centers. All participants were enrolled from January 2018 to October 2018, where they delivered the second baby during this period. A total of 6204 women were enrolled in this study, and 1002 women with a history of GDM were analyzed further. All participants enrolled in the study had an oral glucose tolerance test (OGTT) result at 24 to 28 weeks and were diagnosed as GDM in the first pregnancy according to the OGTT value (when any one of the following values is met or exceeded to the 75-g OGTT: 0 h [fasting], ≥5.10 mmol/L; 1 h, ≥10.00 mmol/L; and 2 h, ≥8.50 mmol/L). The prevalence of GDM recurrence and development of type 2 diabetes mellitus were calculated, and its related risk factors were analyzed. RESULTS: In 6204 participants, there are 1002 women (1002/6204,16.15%) with a history of GDM and 5202 women (5202/6204, 83.85%) without a history of GDM. There are significant differences in age (32.43 ± 4.03 years vs. 33.00 ± 3.34 years vs. 32.19 ± 3.37 years, P  < 0.001), pregnancy interval (4.06 ± 1.44 years vs. 3.52 ± 1.43 years vs. 3.38 ± 1.35 years, P  = 0.004), prepregnancy body mass index (BMI) (27.40 ± 4.62 kg/m2vs. 23.50 ± 3.52 kg/m2vs. 22.55 ± 3.47 kg/m2, P < 0.001), history of delivered macrosomia (22.7% vs. 11.0% vs. 6.2%, P < 0.001) among the development of diabetes mellitus (DM), recurrence of GDM, and normal women. Moreover, it seems so important in the degree of abnormal glucose metabolism in the first pregnancy to the recurrence of GDM and the development of DM. There are significant differences in OGTT levels of the first pregnancy such as area under the curve of OGTT value (18.31 ± 1.90 mmol/L vs. 16.27 ± 1.93 mmol/L vs. 15.55 ± 1.92 mmol/L, P < 0.001), OGTT fasting value (5.43 ± 0.48 mmol/L vs. 5.16 ± 0.49 mmol/L vs. 5.02 ± 0.47 mmol/L, P < 0.001), OGTT 1-hour value (10.93 ± 1.34 mmol/L vs. 9.69 ± 1.53 mmol/L vs. 9.15 ± 1.58 mmol/L, P < 0.001), OGTT 2-hour value (9.30 ± 1.66 mmol/L vs. 8.01 ± 1.32 mmol/L vs. 7.79 ± 1.38 mmol/L, P < 0.001), incidence of impaired fasting glucose (IFG) (fasting plasma glucose ≥5.6 mmol/L) (31.3% vs. 14.6% vs. 8.8%, P < 0.001), and incidence of two or more abnormal OGTT values (68.8% vs. 39.7% vs. 23.9%, P < 0.001) among the three groups. Using multivariate analysis, the factors, such as age (1.07 [1.02-1.12], P = 0.006), prepregnancy BMI (1.07 [1.02, 1.12], P  = 0.003), and area under the curve of OGTT in the first pregnancy (1.14 [1.02, 1.26], P  = 0.02), have an effect on maternal GDM recurrence; the factors, such as age (1.28 [1.01-1.61], P  = 0.04), pre-pregnancy BMI (1.26 [1.04, 1.53], P = 0.02), and area under the curve of OGTT in the first pregnancy (1.65 [1.04, 2.62], P = 0.03), have an effect on maternal DM developed further. CONCLUSIONS The history of GDM was significantly associated with a higher maternal risk for GDM recurrence during follow-up after the first pregnancy. The associated risk factors for GDM recurrence or development of DM include age, high pre-pregnancy BMI, history of delivered macrosomia, the OGTT level in the first pregnancy, such as the high area under the curve of OGTT, IFG, and two or more abnormal OGTT values. To prevent GDM recurrence, women with a history of GDM should do the preconception counseling before preparing next pregnancy.
Adult ; Blood Glucose/metabolism* ; China/epidemiology* ; Diabetes Mellitus, Type 2/epidemiology* ; Diabetes, Gestational ; Female ; Fetal Macrosomia ; Glucose Intolerance ; Humans ; Male ; Pregnancy ; Retrospective Studies

Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection methods.Terminal deoxynucleotidyl transferase(TdT)catalyzes continuous polymerization of deoxynucleotide triphosphates at the 3'-OH end of single-stranded DNA in the absence of a template.In this study,we developed a highly sensitive and selective method based on TdT and endonuclease Ⅳ(Endo Ⅳ)to quantify ALP activity.After ALP hydrolyzes the 3'-PO4 end of the substrate and generates 3'-OH,TdT can effectively elongate the 3'-OH end with deoxynucleotide adenine triphosphate(dATP)and produce a poly A tail,which can be detected by the poly T probes.Endo Ⅳ digests the AP site in poly T probes to generate a fluorescent signal and a new 3'-OH end,leading to the generation of exponential fluorescence signal amplification.The substrate for TdT elongation was optimized,and a limit of detection of 4.3×10-3 U/L was achieved for ALP by the optimized substrate structure.This method can also detect ALP in the cell lysate of a single cell.This work has potential applications in disease diagnosis and biomedical detection.

Objective:To explore the effects of interpregnancy interval (IPI) on pregnancy outcomes of subsequent pregnancy.Methods:A multicenter retrospective study was conducted in 21 hospitals in China. Information of age, height, pre-pregnancy weight, IPI, history of diseases, complications of pregnancy, gestational age of delivery, delivery mode, and pregnancy outcomes of the participants were collected by consulting medical records of pregnant women who had two consecutive deliveries in the same hospital during 2011 to 2018. The participants were divided into 4 groups according to IPI:<18 months, 18-23 months, 24-59 months and ≥60 months. According to the WHO′s recommendation, with the IPI of 24-59 months group as a reference, to the effects of IPI on pregnancy outcomes of subsequent pregnancy were analyzed. Stratified analysis was further carried out based on age, history of gestational diabetes mellitus (GDM), macrosomia, and premature delivery, to explore the differences in the effects of IPI on pregnancy outcomes among women with different characteristics.Results:A total of 8 026 women were included in this study. There were 423, 623, 5 512 and 1 468 participants in <18 months group, 18-23 months group, 24-59 months group and ≥60 months group, respectively. (1) The age, pre-pregnancy body mass index (BMI), history of cesarean section, GDM, gestational hypertension and cesarean section delivery rate of <18 months group, 18-23 months group, 24-59 months group and ≥60 months group were gradually increased, and the differences were statistically significant ( P<0.05). (2) After adjusting for potential confounding factors, compared with women in the IPI of 24-59 months group, the risk of premature delivery, premature rupture of membranes, and oligohydramnios were increased by 42% ( OR=1.42, 95% CI: 1.07-1.88, P=0.015), 46% ( OR=1.46, 95% CI: 1.13-1.88, P=0.004), and 64% ( OR=1.64, 95% CI: 1.13-2.38, P=0.009) respectively for women in the IPI≥60 months group. No effects of IPI on other pregnancy outcomes were found in this study ( P>0.05). (3) After stratified by age and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would significantly increase the risk of oligohydramnios for women with advanced age ( OR=2.87, 95% CI: 1.41-5.83, P=0.004); and <18 months could increase the risk of premature rupture of membranes for women under the age of 35 ( OR=1.59, 95% CI: 1.04-2.43, P=0.032). Both the risk of premature rupture of membranes ( OR=1.58, 95% CI: 1.18-2.13, P=0.002) and premature delivery ( OR=1.52, 95% CI: 1.07-2.17, P=0.020) were significantly increased in the IPI≥60 months group. After stratified by history of GDM and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would lead to an increased risk of postpartum hemorrhage for women with a history of GDM ( OR=5.34, 95% CI: 1.45-19.70, P=0.012) and an increased risk of premature rupture of membranes for women without a history of GDM ( OR=1.44, 95% CI: 1.10-1.90, P=0.009). After stratified by history of macrosomia and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months could increase the proportion of cesarean section for women with a history of macrosomia ( OR=4.11, 95% CI: 1.18-14.27, P=0.026) and the risk of premature rupture of membranes for women without a history of macrosomia ( OR=1.46, 95% CI: 1.12-1.89, P=0.005). After stratified by history of premature delivery and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would significantly increase the risk of premature rupture of membranes for women without a history of premature delivery ( OR=1.47, 95% CI: 1.13-1.92, P=0.004). Conclusions:Both IPI≥60 months and <18 months would increase the risk of adverse pregnancy outcomes in the subsequent pregnancy. Healthcare education and consultation should be conducted for women of reproductive age to maintain an appropriate IPI when they plan to pregnant again, to reduce the risk of adverse pregnancy outcomes in the subsequent pregnancy.

BACKGROUND:Studies have shown that human placenta-derived mesenchymal stem cells with strong proliferative ability have rich sources and can remarkably promote tendon-bone healing after celltransplantation.
OBJECTIVE:To observe the effect of human placenta-derived mesenchymal stem cells on tendon-bone healing in a bone tunnel.
METHODS:Human placenta-derived mesenchymal stem cells were separated using adherent separation screening method. Thirty 8-week-old male Sprague-Dawley rats were randomly divided into two groups, 15 rats as experimental group and 15 rats as control group. Experimental group were subjected to transplantation of human placenta-derived mesenchymal stem cells and the control group were injected with saline solution.
RESULTS AND CONCLUSION:Stem cells, new vessels, and fibrocartilage hyperplasia were observed on the tendon-bone interface with microscope at 2, 4 and 6 weeks after celltransplantation in the experimental group. Biomechanical y, the maximum pul out load in the experimental group was significantly higher than that in the control group at 4 and 6 weeks after celltransplantation (P<0.05). These findings suggest that human placenta-derived mesenchymal stem cells can accelerate early tendon-bone healing in a bone tunnel and strengthen the biomechanical strength.