Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

OBJECTIVES: Liver disease is the most common extra-intestinal manifestation of ulcerative colitis (UC), but the underlying pathogenesis is still not clarified. It is well accepted that the occurrence of UC-related liver disease has close correlation with immune activation, intestinal bacterial liver translocation, inflammatory cytokine storm, and the disturbance of bile acid circulation. The occurrence of UC-related liver disease makes the therapy difficult, therefor study on the pathogenesis of UC-related liver injury is of great significance for its prevention and treatment. Glutathione (GSH) shows multiple physiological activities, such as free radical scavenging, detoxification metabolism and immune defense. The synthesis and the oxidation-reduction all contribute to GSH antioxidant function. It is reported that the deficiency in hepatic GSH antioxidant function participates in multiple liver diseases, but whether it participates in the pathogenesis of UC-related liver injury is still not clear. This study aims to investigate the feature and underlying mechanism of GSH synthesis and oxidation-reduction function during the development of UC, which will provide useful information for the pathogenesis study on UC-related liver injury. METHODS: UC model was induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS)-ethanol solution (5 mg/0.8 mL per rat, 50% ethanol) via intra-colonic administration in rats, and the samples of serum, liver, and colon tissue of rats were collected at the 3rd, 5th, and 7th days post TNBS. The severity degree of colitis was evaluated by measuring the disease activity index, colonic myeloperoxidase activity, and histopathological score, and the degree of liver injury was evaluated by histopathological score and the serum content of alanine aminotransferase. Spearman correlation analysis was also conducted between the degree of colonic lesions and index of hepatic histopathological score as well as serum aspartate aminotransferase level to clarify the correlation between liver injury and colitis. To evaluate the hepatic antioxidant function of GSH in UC rats, hepatic GSH content, enzyme activity of GSH peroxidase (GSH-Px), and GSH reductase (GR) were determined in rats at the 3rd, 5th, and 7th days post TNBS, and the protein expressions of glutamine cysteine ligase (GCL), GSH synthase, GSH-Px, and GR in the liver of UC rats were also examined by Western blotting. RESULTS: Compared with the control, the disease activity index, colonic myeloperoxidase activity, and histopathological score were all significantly increased at the 3rd, 5th, and 7th days post TNBS (all P<0.01), the serum aspartate aminotransferase level and hepatic histopathologic score were also obviously elevated at the 7th day post TNBS (all P<0.05). There was a significant positive correlation between the degree of liver injury and the severity of colonic lesions (P=0.000 1). Moreover, compared with the control, hepatic GSH content and the activity of GSH-Px and GR were all significantly decreased at the 3rd and 5th days post TNBS (P<0.05 or P<0.01), and the protein expressions of GCL, GSH-Px, and GR were all obviously down-regulated at the 3rd, 5th, and 7th days post TNBS (P<0.05 or P<0.01). CONCLUSIONS There is a significant positive correlation between the degree of liver injury and the severity of colonic lesions, and the occurrence of reduced hepatic GSH synthesis and decreased GSH reduction function is obviously earlier than that of the liver injury in UC rats. The reduced hepatic expression of enzymes that responsible for GSH synthesis and reduction may contribute to the deficiency of GSH synthesis and oxidation-reduction function, indicating that the deficiency in GSH antioxidant function may participate in the pathogenesis of UC related liver injury.
Animals ; Antioxidants ; Aspartate Aminotransferases ; Colitis/chemically induced* ; Colitis, Ulcerative/metabolism* ; Colon/pathology* ; Glutathione/biosynthesis* ; Liver/metabolism* ; Peroxidase/metabolism* ; Rats ; Trinitrobenzenesulfonic Acid

Objective:To establish the normal values of water-perfused high resolution esophageal manometry (HREM)(GAP-36A) at resting period, water swallowing, semisolid swallowing and solid swallowing in Chinese population.Methods:From September 1, 2019 to June 30, 2020, 91 healthy volunteers receiving water-perfused HREM (GAP-36A) at resting period, water swallowing, semisolid swallowing and solid swallowing were selected from 9 hospitals (Union Hospital, Tongji Medical College, Huazhong University of Science and Technology; the First Affiliated Hospital of Dalian Medical University; the Second Hospital of Hebei Medical University; the Second Affiliated Hospital, Naval Medical University; the First Affiliated Hospital, Sun Yat-sen University; the First Affiliated Hospital, University of Science and Technology of China; Aviation General Hospital of China Medical University; the Affiliated Hospital of Medical School of Nanjing University and the First People′s Hospital of Yichang). Parameters included the position of the upper and lower edges of the upper esophageal sphincter (UES) and lower esophageal sphincter (LES), the length of the LES and UES, the position of the pressure inversion point (PIP), the resting pressure of UES and LES and swallow-related parameters such as the distal contraction integral (DCI), 4 s integrated relaxation pressure (IRP), distal latency (DL) and UES residual pressure. One-way analysis of variance, post-hoc test and sum rank test were used for statistical analysis.Results:A total of 87 healthy volunteers were enrolled, including 40 males and 47 females, aged (38.5±14.2) years old (ranged from 19 to 65 years old). The position of the upper and lower edges of the LES was (42.7±2.8) and (45.6±2.8) cm, respectively, the length of the LES was (2.9±0.4) cm, and the position of PIP was (43.3±2.8) cm. The position of the upper and lower edges of the UES was (18.1±3.0) and (22.6±2.0) cm, respectively, and the length of the UES was (4.8±1.0) cm. The resting pressure of LES and UES was (17.4±10.7) and (84.1±61.1) mmHg (1 mmHg=0.133 kPa), respectively. The DCI value at solid swallowing was higher than those at water swallowing and semisolid swallowing ((2 512.4±1 448.0) mmHg·s·cm vs. (2 183.2±1 441.2) and (2 150.8±1 244.8) mmHg·s·cm), and the differences were statistically significant ( t=-4.30 and -3.74, both P<0.001). The values of 4 s IRP at semisolid swallowing and solid swallowing were lower than that at water swallowing ((4.6±4.1) and (4.9±3.9) mmHg vs. (5.4±3.9) mmHg), and the differences were statistically significant ( t=3.38 and 2.09, P=0.001 and 0.037). The DL at water swallowing was shorter than those at semisolid swallowing and solid swallowing ((8.5±1.8) s vs. (9.8±2.2) and (10.6±2.8) s), and the DL at semisolid swallowing was shorter than that at solid swallowing, and the differences were statistically significant ( t=-10.21, -13.91 and -4.68, all P<0.001). The UES residual pressure at water swallowing was higher than those at semisolid swallowing and solid swallowing (9.5 mmHg, 6.5 to 12.3 mmHg vs. 8.0 mmHg, 4.5 to 11.7 mmHg and 5.5 mmHg, 2.0 to 9.3 mmHg), and the UES residual pressure at semisolid swallowing was higher than that at solid swallowing, and the differences were statistically significant ( t=3.48, 10.30 and 6.35, all P<0.001). Conclusions:The normal values of water-perfused HREM (GAP-36A) in Chinese population at resting period, water swallowing, semisolid swallowing and solid swallowing can provide a reference basis for clinical diagnosis and treatment for patients receiving water-perfused HREM examination.

Objective:To investigate the safety and efficacy of sirolimus through the change in platelet counts during the sirolimus therapy on Kasaback-Merritt phenomenon(KMP).Methods:Four patients were treated in Nanjing Children Hospital between Jaunary 2017 and June 2019 were enrolled in the study, including two males and two females. Their age was ranged from two days to three months. They all presented with huge mass located in neck or retroperitoneal and thrombocytopenia, hypofibrinogenemia. Their coagulation could not be improved by surgery or palate transfusion or steroids. They were all diagnosed as Kaposiform hemangioendothelioma(KHE) with KMP according to the biopsy, coagulation index and CT. Sirolimus were administered with 0.1 mg -1·kg -1·d -1 or 0.8 mg/m 2, twice daily. Subsequent dose was adjusted to maintain the trough level between 10-15 ng/ml.Steroids were weaned gradually. Blood accounts were measured following the sirolimus administration. Results:All the four patients got improvement in platelet counts. Three patients went through a significant decreasing of palate counts after adding sirolimus for about 1-4 days. Two lesions prompted improvement with smaller size, lighter color and softer texture without complete regression. Two patients had elevated liver enzymes and/or interstitial pneumonia. One recovered while the other died of severe pneumonia with dyspnea. All the palate counts of survived patients remained in normal level in the following one year.Conclusions:Sirolimus is an effective method in treating KMP and stabilizing platelet counts. However, it can not cure hemangioendothelioma.

Objective:To investigate the safety and efficacy of sirolimus through the change in platelet counts during the sirolimus therapy on Kasaback-Merritt phenomenon(KMP).Methods:Four patients were treated in Nanjing Children Hospital between Jaunary 2017 and June 2019 were enrolled in the study, including two males and two females. Their age was ranged from two days to three months. They all presented with huge mass located in neck or retroperitoneal and thrombocytopenia, hypofibrinogenemia. Their coagulation could not be improved by surgery or palate transfusion or steroids. They were all diagnosed as Kaposiform hemangioendothelioma(KHE) with KMP according to the biopsy, coagulation index and CT. Sirolimus were administered with 0.1 mg -1·kg -1·d -1 or 0.8 mg/m 2, twice daily. Subsequent dose was adjusted to maintain the trough level between 10-15 ng/ml.Steroids were weaned gradually. Blood accounts were measured following the sirolimus administration. Results:All the four patients got improvement in platelet counts. Three patients went through a significant decreasing of palate counts after adding sirolimus for about 1-4 days. Two lesions prompted improvement with smaller size, lighter color and softer texture without complete regression. Two patients had elevated liver enzymes and/or interstitial pneumonia. One recovered while the other died of severe pneumonia with dyspnea. All the palate counts of survived patients remained in normal level in the following one year.Conclusions:Sirolimus is an effective method in treating KMP and stabilizing platelet counts. However, it can not cure hemangioendothelioma.

Background and Objectives: Cranial sutures play a critical role in adjustment of skull development and brain growth. Premature fusion of cranial sutures leads to craniosynostosis. The aim of the current study was to culture and characterize human cranial suture mesenchymal cells in vitro. Methods: The residual skull tissues, containing synostosed or contralateral suture from three boys with right coronal suture synostosis, were used to isolate the suture mesenchymal cells. Then, flow cytometry and multilineage differentiation were performed to identify the typical mesenchymal stem cell (MSC) properties. Finally, we used quantitative real-time polymerase chain reaction (RT-PCR) to detect the mRNA expression of osteogenesis and stemness related genes. Results: After 3 to 5 days in culture, the cells migrated from the tissue explants and proliferated parallelly or spirally. These cells expressed typical MSC markers, CD73, CD90, CD105, and could give rises to osteocytes, adipocytes and chondrocytes. RT-PCR showed relatively higher levels of Runx2, osteocalcin and FGF2 in the fused suture MSCs than in the normal cells. However, BMP3, the only protein of BMP family that inhibits osteogenesis, reduced in synostosed suture derived cells. The expression of effector genes remaining cell stemness, including Bmi1, Gli1 and Axin2, decreased in the cells migrated from the affected cranial sutures. Conclusions The MSCs from prematurely occlusive sutures overexpressed osteogenic related genes and down-regulated stemness-related genes, which may further accelerate the osteogenic differentiation and suppress the self-renewal of stem cells leading to craniosynostosis.

Objective: To explore the effects of Bmi1 on proliferation of mouse cranial suture mesenchymal cells. Methods: Primary posterior frontal and sagittal suture derived cells were isolated from the 2-5 d old C57BL/6 suckling mice (n=6) of the same brood and cultured. Flow cytometry and multilineage differentiation assay were performed to identify the mesenchymal stem cells (MSCs) characteristics of the 2 kinds of cranial suture-derived cells. The mRNA expression of stem cell related genes, Bmi1, Twist1, Gli1 and Axin2 were detected by real-time quantitative polymerase chain reaction (RT-PCR). Then, the proliferation and downstream protein expression were analyzed after down-regulation of Bmi1 in the sagittal suture derived MSCs by transfecting Bmi1 siRNA. The t test was used to compare the mean between two groups. Statistical significance was set at P<0.05. Results: The mouse cranial suture derived cells were successfully cultured in vitro. These cells expressed typical MSCs markers, CD44, CD90, CD73, except for CD34. These cells had osteogenic, adipogenic and chondrogenic differentiation potency. RT-PCR results showed that the mRNA expressions of Bmi1 (0.006 30±0.000 58 vs 0.002 60±0.000 34, t=5.430, P=0.005 6), Twist1(0.000 31±0.000 04 vs 0.000 15±0.000 02, t=3.343, P=0.028 8), Axin2(0.000 33±0.000 03 vs 0.000 17±0.000 05, t=3.067, P=0.037 4) and Gli1 (0.001 10±0.000 13 vs 0.000 60±0.000 33, t=3.956, P=0.016 7) were significantly decreased in the posterior frontal suture MSCs compared with those in sagittal suture derived cells. Among them, Bmi1 has the largest decline. After down-regulation of Bmi1 in sagittal suture MSCs, the protein expression level of Ink4a was significantly up-regulated compared with the control group, and the cell proliferation ability was significantly decreased. Conclusions Inhibition of Bmi1 expression can up-regulate the expression of Ink4a protein and decrease the proliferation ability of suture MSCs, which may lead to craniosynostosis.

Objective: To observe the dynamic changes of plasma level thymosinβ4 (Tβ4) in acute myocardial infarction (AMI) patients with intervening therapy within 15 days of onset and to explore the relationship between Tβ4 and clinical prognosis in AMI patients.
Methods: Our research included 2 groups:AMI group, n=69 and Control group, the patients with suspected chest pain while CAG excluded coronary artery stenosis, n=32. Plasma levels of Tβ4 were examined in all AMI patients on admission day and every day until 15 days of onset;AMI patients were followed-up for 18 months and the endpoint was defined as major adverse cardiovascular event (MACE) occurrence.
Results: ①Compared with Control group, AMI group had increased plasma level of Tβ4 on admission day and on day-15 of onset, P<0.01. ② With intervening therapy, AMI group had elevated Tβ4 level upon immediate onset, it was decreased on day-1, reached low level on day-3 and elevated to peak on day-6, then reduced followed by slightly raising on day-11.③During follow-up period, the AMI patients without MACE had the higher mean in-hospital maximum Tβ4 value than those with MACE occurrence, P<0.01. Logistic regression analysis indicated that the mean in-hospital maximum Tβ4 value was related to MACE occurrence during follow-up period (OR=0.999, 95%CI 0.999-1.000).
Conclusion: AMI may induce up-regulated expression of plasma Tβ4;with intervening therapy, Tβ4 showed a trend of“elevation-reduction-elevation-reduction”at the early stage of AMI. High expression of Tβ4 was helpful for improving clinical prognosis in AMI patients which may provide a theoretical basis for exogenous use of Tβ4 in AMI treatment.

Objective To investigate the regulatory effects of cyclic diguanylate (c-di-GMP) signaling on CheB and CheR, which were chemotaxis regulatory proteins relating to the motility of Leptospira interrogans.Methods Real-time PCR was used to determine the expression of cheB1, cheB2, cheB3, cheR1 and cheR2 genes at mRNA level during Leptospira interrogans infection.Fragments of these genes were amplified and cloned into the expression vector pET-28a, respectively, to construct the prokaryotic expression system for them.Colony morphologies of Escherichia coli (E.coli) strains that overexpressed the target genes were observed to determine the regulatory effects of c-di-GMP on CheB and CheR.Results The expression of cheB1 gene at mRNA level increased 60 min after infection and reached the peak at 90 min.Compared with the control group, the expression of cheB3 gene at mRNA level were up-regulated, while no significant difference in the expression of cheB2 and cheR genes was observed 60 min after infection.The prokaryotic expression system for the five genes was successfully constructed and the purified proteins were obtained.CheB1, CheB3 and CheR2 improved the motility of E.coli, but that was inhibited by the inhibitor of diguanylate cyclase (DGC) or phosphodiesterase (PDE).Conclusion CheB and CheR regulate the swarming motility of E.coli and are affected by intracellular c-di-GMP.

This paper reported 13 cases of delayed traumatic chest wall abscess from January 2012 to January 2015.All the patients were associated with type 2 diabetes.After local puncture for confirmative diagnosis, a chest wall abscess dissection was carried out as soon as possible.At each site of upper and lower pole, an indwelling drainage tube was placed for irrigation and negative pressure suction.Sensitive antibiotics were selected based on susceptibility test results.The drainage tubes were removed 7-14 days after surgery.There were 8 cases of primary healing of incision and 5 cases of secondary healing of incision.All the patients were cured.Follow-ups for 6-36 months (average, 17 months) showed no recurrence.