Chronic visceral pain is a persistent and debilitating condition arising from dysfunction or sensitization of the visceral organs and their associated nervous pathways. Increasing evidence suggests that imbalances in central nervous system function play an essential role in the progression of visceral pain, but the exact mechanisms underlying the neural circuitry and molecular targets remain largely unexplored. In the present study, the ventral tegmental area (VTA) was shown to mediate visceral pain in mice. Visceral pain stimulation increased c-Fos expression and Ca²⁺ activity of glutamatergic VTA neurons, and optogenetic modulation of glutamatergic VTA neurons altered visceral pain. In particular, the upregulation of NMDA receptor 2A (NR2A) subunits within the VTA resulted in visceral pain in mice. Administration of a selective NR2A inhibitor decreased the number of visceral pain-induced c-Fos positive neurons and attenuated visceral pain. Pharmacology combined with chemogenetics further demonstrated that glutamatergic VTA neurons regulated visceral pain behaviors based on NR2A. In summary, our findings demonstrated that the upregulation of NR2A in glutamatergic VTA neurons plays a critical role in visceral pain. These insights provide a foundation for further comprehension of the neural circuits and molecular targets involved in chronic visceral pain and may pave the way for targeted therapies in chronic visceral pain.
Animals ; Male ; Visceral Pain/metabolism* ; Up-Regulation/physiology* ; Ventral Tegmental Area/metabolism* ; Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors* ; Neurons/drug effects* ; Mice, Inbred C57BL ; Mice ; Proto-Oncogene Proteins c-fos/metabolism* ; Chronic Pain/metabolism* ; Glutamic Acid/metabolism*

Objective:To investigate the value of 3D CT reconstruction in diagnosis of chronic lateral ankle instability (CLAI) combined with syndesmotic diastasis (SD).Methods:A retrospective study was conducted to analyze the clinical data of 160 patients with CLAI who had been examined by arthroscopy from January 2018 to September 2022 at Department of Foot and Ankle Surgery, Wuhan Fourth Hospital. There were 64 males and 96 females with an age of (39.8±12.6) years. Eighty-one left and 79 right feet were affected; the time from injury to surgery was (27.3±11.6) months. The patients were divided into a widened interval group and a normal interval group according to the syndesmotic width measured, with 2 mm as a critical value. After preoperative 3D CT reconstruction, the differences in anterior tibiofibular distance, posterior tibiofibular distance, the narrowest tibiofibular distance, fibular translation, fibular rotation, and syndesmotic area (SA) were compared between the 2 groups. Univariate and multivariate analyses were performed successively to identify the risk factors. The receiver operating characteristic (ROC) curve was used to identify the best predictive factor and critical value. According to the findings of previous research, the above analyses were repeated to determine the best predictive factor and critical value respectively in the sex subgroup, fibular morphology subgroup and incisura feature subgroup.Results:The binary logistic regression showed that SA was a risk factor for CLAI combined with SD ( OR=1.196, 95% CI: 1.122 to 1.275, P < 0.001). The ROC curve revealed an area under curve of 0.847 and the difference critical value of 22.06 mm 2 that indicated a sensitivity of 80.4% and a specificity of 78.9%, respectively. Subgroup analyses showed that SA was suitable for male and female patients and patients with different fibular morphologies and incisura features but the difference critical values were different. Conclusion:In 3D CT reconstruction, measurement of SA may help the diagnosis of CLAI combined with SD.

Objective To analyze risk factors of acute respiratory failure(ARF)in patients with hypertriglyceridemia acute pancreatitis(HTG-AP)and construct a risk prediction model.Methods A total of 222 HTG-AP patients were included in this study and divided into the non-ARF group(176 cases)and the ARF group(46 cases)according to diagnostic guidelines for ARF.Clinical data of the two groups were compared and the predictive factors were screened.These selected factors were then utilized in a multivariate Logistic regression analysis to construct a Logistic regression model.Subsequent evaluation of the model′s predictive ability,accuracy and clinical utility was conducted through ROC,curve analysis,calibration plot examination and decision curve analysis(DCA),respectively.Results Compared with the non-ARF group,the levels of high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL)-C and albumin(ALB)were decreased in the ARF group(P＜0.05),while the levels of creatinine(Cr),urea nitrogen(BUN),aspartate aminotransferase(AST)and C-reactive protein(CRP)were increased,and the incidence of pleural fluid and ascites was also increased(P＜0.05).Multivariate Logistic regression analysis showed that higher levels of Cr and AST,lower levels of ALB,HDL-C and ascites were independent risk factors for HTG-AP complicated ARF(P＜0.05).Based on these results,a column-line prediction model for HTG-AP complicated ARF was established.After internal verification,the area under curve(AUC)of receiver operating characteristic(ROC)curve of the nomogram model was 0.952(95%CI:0.923-0.981),the Youden index was 0.808 and the sensitivity and specificity were 93.33%and 87.43%,respectively.The calibration curve showed that the probability of HTG-AP concurrent ARF predicted by the model was in good agreement with the actual probability.The DCA curve showed that the model had certain clinical value.Conclusion The nomogram prediction model combined could provide a scheme for the clinical prevention of HTG-AP complicated with ARF.

OBJECTIVE To study the effects of bergapten in the treatment of liver fibrosis and its mechanism based on serum metabolomics. METHODS Forty mice were divided into normal control group （0.5% carboxymethyl cellulose sodium solution）， model group （0.5% carboxymethyl cellulose sodium solution）， and BP low-dose and high-dose groups （50， 100 mg/kg）， with 10 mice in each group. Except for the normal control group， the other three groups were all treated with carbon tetrachloride to induce liver fibrosis model； they were given relevant medicine/solution intragastrically， once a day， for consecutive 8 weeks. After the last medication， the levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum were detected， and liver pathological changes were observed； the expressions of α-smooth muscle actin （α-SMA） and Collagen Ⅰ were detected in liver tissue； the serum of the mice was collected for metabolomics analysis. RESULTS Compared with the model group， serum levels of ALT and AST and protein expressions of α-SMA and Collagen Ⅰ in liver tissue were decreased significantly in BP high-dose and low-dose groups （P＜0.05）， while liver fibrosis was improved significantly. Meanwhile， metabolomics analyses showed that there were a total of 175 serum differential metabolites in the BP high-dose group and model group， of which 18 substances were upregulated and 157 substances were downregulated； the main metabolic pathways involved in bergapten intervention were pyrimidine metabolism， butanoate metabolism， fatty acid synthesis， tyrosine metabolism， β-alanine metabolism， nicotinic acid and nicotinamide metabolism， glutathione metabolism， etc. CONCLUSIONS BP is effective in the treatment of liver fibrosis by regulating pyrimidine metabolism， butanoate metabolism， glutathione metabolism and so on in rats with liver fibrosis.

Objective:To explore the clinical characteristics and distribution of Tranditional Chinese Medicine (TCM) syndrome types of refractory tic disorders in children based on clustering analysis.Methods:A cross-sectional study. 183 children aged 3-18 years with refractory tic disorders from pediatric encephalopathy outpatient department of the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine who met the inclusion criteria from October 2015 to January 2022 were recruited. Frequency descriptive analysis, systematic clustering analysis and principal component analysis were conducted by Microsoft Office Excel 2016 and IBM SPSS Statistics 26.0 to explore the clinical characteristics and syndrome distribution of the disease.Results:The TCM symptoms of 183 children with refractory tic disorders were clustered into 6 categories, and finally 5 types of syndrome were obtained, among which the syndrome of spleen deficiency and liver hyperactivity mixed with dampness accounted for the largest proportion (27.32%), followed by the syndrome of phlegm-fire disturbing spirit (21.31%), the syndrome of wind-heat invading lung (18.03%), the syndrome of phlegm-qi stagnation (17.49%), and the syndrome of dampness and yin deficiency (15.85%).Conclusion:Through the mining and analysis of the outpatient information of pediatric encephalopathy in our hospital, the common syndrome differentiation types of refractory tic disorder are obtained, which can provide a reference for the TCM syndrome differentiation types of refractory tic disorder in children.

Periodontitis is caused by overactive osteoclast activity that results in the loss of periodontal supporting tissue and mesenchymal stem cells (MSCs) are essential for periodontal regeneration. However, the hypoxic periodontal microenvironment during periodontitis induces the apoptosis of MSCs. Apoptotic bodies (ABs) are the major product of apoptotic cells and have been attracting increased attention as potential mediators for periodontitis treatment, thus we investigated the effects of ABs derived from MSCs on periodontitis. MSCs were derived from bone marrows of mice and were cultured under hypoxic conditions for 72 h, after which ABs were isolated from the culture supernatant using a multi-filtration system. The results demonstrate that ABs derived from MSCs inhibited osteoclast differentiation and alveolar bone resorption. miRNA array analysis showed that miR-223-3p is highly enriched in those ABs and is critical for their therapeutic effects. Targetscan and luciferase activity results confirmed that Itgb1 is targeted by miR-223-3p, which interferes with the function of osteoclasts. Additionally, DC-STAMP is a key regulator that mediates membrane infusion. ABs and pre-osteoclasts expressed high levels of DC-STAMP on their membranes, which mediates the engulfment of ABs by pre-osteoclasts. ABs with knock-down of DC-STAMP failed to be engulfed by pre-osteoclasts. Collectively, MSC-derived ABs are targeted to be engulfed by pre-osteoclasts via DC-STAMP, which rescued alveolar bone loss by transferring miR-223-3p to osteoclasts, which in turn led to the attenuation of their differentiation and bone resorption. These results suggest that MSC-derived ABs are promising therapeutic agents for the treatment of periodontitis.
Humans ; Osteoclasts ; Alveolar Bone Loss/therapy* ; Cell Differentiation ; MicroRNAs ; Periodontitis/therapy* ; Extracellular Vesicles ; Apoptosis ; Mesenchymal Stem Cells

Objective:To study the treatment outcomes of transjugular intrahepatic portal shunt (TIPS) on refractory hepatic sinus obstruction syndrome (HSOS) caused by Gynura segetum.Methods:The clinical data of 15 patients with refractory HSOS caused by Gynura segetum treated at the Department of Vascular Surgery, Henan Provincial People's Hospital from January 2017 to April 2021 were retrospectively analyzed. There were 7 males and 8 females, with ages ranging from 30 to 85 years, mean ± s. d. (61.2±14.1) years. Albumin, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, glutamyl transferase, and portal vein pressure were compared before and after TIPS. The liver function and renal function of these patients were followed up.Results:When compared with pre-operation, the albumin, alanine aminotransferase, aspartate aminotransferase and other indexes were significantly improved after TIPS (all P<0.05). The portal vein pressure of 15 patients significantly decreased from the preoperative volume of (41.7±3.5) cmH 2O (1 cmH 2O=0.098 kPa) to (28.3±4.4) cmH 2O ( t=10.41, P<0.001). The preoperative liver function was Child-Pugh grade A in 1 patient, grade B in 8 patients, grade C in 6 patients. The postoperative Child-Pugh grading was grade A in 14 patients and grade B in 1 patient. Ascites, gastrointestinal bleeding, abdominal pain, abdominal distention and spontaneous peritonitis all disappeared in these 15 patients. Postoperative hepatic encephalopathy developed in 2 patients and hepatic myelopathy in 1 patient. Conclusion:TIPS for treatment of HSOS caused by Gynura segetum resulted in a rapid recovery of liver function, rapid symptomatic relief, with a low incidence of hepatic encephalopathy/hepatic myelopathy.

Objective:To investigate the surgical treatment, clinical effect and revision reasons of children with proximal femoral fibrous dysplasia(FD).Methods:The clinical data of 26 children with polyostotic FD of proximal femur who underwent surgery at Department of Pediatric Orthopaedics, Beijing Jishuitan Hospital from June 2016 to June 2018 were retrospectively analyzed. There were 18 males and 8 females with a mean age of 9.2 years (range:5 to 16 years).One of them was McCune Albright syndrome. Fifteen cases were in first operation and 11 cases were in revision operation. The operation methods and results were reviewed,and the causes of revision were analyzed.Results:Among the 15 children who underwent the first operation,13 cases underwent osteotomy or fracture reduction and interlocking intramedullary nail(IMN) fixation;One case underwent valgus osteotomy and pediatric hip plate(PHP)internal fixation;One case underwent valgus osteotomy+lesion curettage+allogeneic bone graft+PHP fixation. Among the 11 children who underwent revision surgery,9 cases were treated with IMN fixation,1 case with PHP fixation,and 1 case with PHP fixation+allogeneic bone graft. The causes of revision included distal fixation failed in 6 cases,proximal fixation failed in 3 cases,plate fixation failed in 5 cases,and recurrence occurred after curettage and artificial bone graft in 2 cases. Patients were followed up for 1.4 years(range:1.0 to 3.5 years) after recent operation. The osteotomy or fracture healed well with good deformity correction. Postoperative complications included infection in 1 case and local bone partial resorption in 1 case.Conclusions:Osteotomy combined with rigid internal fixation is an effective surgical treatment for fibrous dysplasia of proximal femur in children. Internal fixation should cover the whole length of lesion. Intramedullary nail is the most common choice. Because the growth of height and the progress of the disease itself,this deformity is prone to recur in children,needing closely follow-up after operation.

Objective:To investigate the surgical treatment, clinical effect and revision reasons of children with proximal femoral fibrous dysplasia(FD).Methods:The clinical data of 26 children with polyostotic FD of proximal femur who underwent surgery at Department of Pediatric Orthopaedics, Beijing Jishuitan Hospital from June 2016 to June 2018 were retrospectively analyzed. There were 18 males and 8 females with a mean age of 9.2 years (range:5 to 16 years).One of them was McCune Albright syndrome. Fifteen cases were in first operation and 11 cases were in revision operation. The operation methods and results were reviewed,and the causes of revision were analyzed.Results:Among the 15 children who underwent the first operation,13 cases underwent osteotomy or fracture reduction and interlocking intramedullary nail(IMN) fixation;One case underwent valgus osteotomy and pediatric hip plate(PHP)internal fixation;One case underwent valgus osteotomy+lesion curettage+allogeneic bone graft+PHP fixation. Among the 11 children who underwent revision surgery,9 cases were treated with IMN fixation,1 case with PHP fixation,and 1 case with PHP fixation+allogeneic bone graft. The causes of revision included distal fixation failed in 6 cases,proximal fixation failed in 3 cases,plate fixation failed in 5 cases,and recurrence occurred after curettage and artificial bone graft in 2 cases. Patients were followed up for 1.4 years(range:1.0 to 3.5 years) after recent operation. The osteotomy or fracture healed well with good deformity correction. Postoperative complications included infection in 1 case and local bone partial resorption in 1 case.Conclusions:Osteotomy combined with rigid internal fixation is an effective surgical treatment for fibrous dysplasia of proximal femur in children. Internal fixation should cover the whole length of lesion. Intramedullary nail is the most common choice. Because the growth of height and the progress of the disease itself,this deformity is prone to recur in children,needing closely follow-up after operation.

Objective:To investigate the effects of tantalum coating on adhesion, proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods:In this study, BMSCs were extracted from 6 6-week-old rats and cultured in vitro to the third generation. Tantalum coating was manufactured on Ti6Al4V by chemical vapor deposition. The cells were identified by flow cytometry before they were induced with different mediums for osteogenesis, chondrogenesis and adipogenesis. The adhesion, proliferation and osteogenic differentiation of BMSCs were detected with fluorescence staining, Cell Counting Kit-8 (CCK8) assay and Q-PCR, respectively. Recorded and compared were the adhesion rate, proliferation rate, and expression of osterix (OSX), Runt-related transcription factor 2 (RUNX2), osteonectin (OSN) and osteopontin (OPN) of BMSCs on the surface of titanium alloy round plates (the Ti6Al4V group) and of tantalum coating round plates (the Ta group). Results:The flow cytometry revealed CD44 (94.55%), CD90 (95.01%) and CD34 (0.06%). Alkaline phosphatase (ALP) staining was positive after osteogenic induction for 14 days; Alizarin red staining showed calcified nodules after osteogenic induction for 21 days; oil red O staining was positive after adipogenic induction for 21 days; alcian blue staining found chondrogenic ability after chondrogenic induction for 21 days. Laser confocal microscopy showed that the BMSCs grew in patches aggregated and closely linked on the surface of titanium alloy round plates (in the Ti6Al4V group) and of tantalum coating round plates (in the Ta group). More BMSCs adhered on the tantalum coating plates than on the titanium alloy plates and exhibited better ductility. The proliferation rates of BMSCs on tantalum coating were significantly faster than those on titanium alloy after 1, 3, 5 and 7 days of co-culture in vitro ( P<0.05).Q-PCR showed that tantalum coating promoted the expression of OSN and OPN after 7 days of culture significantly higher than titanium alloy did ( P<0.05).After 21 days of co-culture in vitro, tantalum coating enhanced the expression of OSX, RUNX2, OSN and OPN significantly higher than titanium alloy did ( P<0.05). Conclusion:Compared to titanium alloy which is used for conventional orthopedic implants, tantalum coating can observably promote adhesion, proliferation and osteogenic differentiation of BMSCs.