ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction （枳壳甘草汤， ZGD） delays intervertebral disc degeneration （IDD） based on the Hippo-yes-associated protein （YAP）/transcriptional co-activator with PDZ-binding motif （TAZ） signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group， model group， low-dose ZGD group， high-dose ZGD group， and high-dose ZGD + inhibitor group， with 10 rats in each group. In the sham surgery group， the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle （depth approximately 2 mm） without damaging the intervertebral disc. In the other groups， rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling， rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/（kg·d） of the decoction via gastric gavage， respectively. The high-dose ZGD + inhibitor group was given 12.24 g/（kg·d） of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/（kg·d） of normal saline via gavage. The gavage was given once a day， and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention， the pathological changes of the tail intervertebral discs were observed using HE staining， Oil Red O-Green staining， and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus， and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway， including YAP， phosphorylated YAP （p-YAP）， phosphorylated MST1/2 （p-MST1/2）， phosphorylated TAZ （p-TAZ） and apoptosis-related proteins， such as Cleaved Caspase 3， P53， Bcl-2 and Bax. ResultsCompared with sham surgery group， the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan， Bcl-2， and YAP proteins in the nucleus pulposus decreased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate increased （P < 0.01）. Compared with the model group， the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan， Bcl-2， and YAP proteins increased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate decreased （P＜0.05 or P＜0.01）. The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group， with a decrease in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and an increase in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. Compared with the high-dose ZGD group， the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration， with an increase in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and a decrease in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus， maintaining the function of the Hippo-YAP/TAZ signaling pathway， and reducing apoptosis of nucleus pulposus cells.

In 2025， Severe Liver Disease and Artificial Liver Group and Nutrition and Regeneration in End-Stage Liver Disease Group of Chinese Society of Hepatology， Chinese Medical Association， convened a panel of national experts to jointly develop China’s first guideline for the diagnosis and treatment of acute-on-chronic liver failure （ACLF）. Based on the latest research findings and clinical practice in China and globally， this guideline establishes a standardized definition of ACLF and provide recommendations for its diagnosis， treatment， and clinical management. This article gives an interpretation of the key points in the guideline， in order to provide a reference for standardized diagnosis and treatment of ACLF.

ObjectiveTo establish and validate a new prediction model for the risk of death in patients with acute-on-chronic liver failure （ACLF） based on sarcopenia and other clinical indicators， and to improve the accuracy of prognostic assessment for ACLF patients. MethodsA total of 380 patients with ACLF who were admitted to Beijing YouAn Hospital， Capital Medical University， from January 2019 to January 2022 were enrolled， and they were divided into training group with 228 patients and testing group with 152 patients in a ratio of 6∶4 using the stratified random sampling method. For the training group， CT images were used to measure the cross-sectional area of the skeletal muscle at the third lumbar vertebra （L3）， and L3 skeletal muscle index （L3-SMI） was calculated. Sarcopenia was diagnosed based on the previously established L3-SMI reference values for healthy adults in northern China. Univariate and multivariable Cox regression analyses were used to establish a sarcopenia-ACLF model which integrated sarcopenia and clinical risk factors， and a nomogram was developed for presentation. The area under the ROC curve （AUC） was used to assess the predictive performance of the model， the calibration curve was used to assess the degree of calibration， and a decision curve analysis was used to investigate the clinical application value of the model. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups， and the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier method was used to plot survival curves， and the Log-rank test was used for comparison between groups. The DeLong test was used for comparison of AUC between different models. ResultsThe multivariate Cox regression analysis showed that sarcopenia （hazard ratio ［HR］=1.962， 95% confidence interval ［CI］： 1.185‍ ‍—‍ ‍3.250， P=0.009）， total bilirubin （HR=1.003， 95%CI： 1.002‍ ‍—‍ ‍1.005， P<0.001）， international normalized ratio （HR=1.997， 95%CI： 1.674‍ ‍—‍ ‍2.382， P<0.001）， and lactic acid （HR=1.382， 95%CI： 1.170‍ ‍—‍ ‍1.632， P<0.001） were included in the sarcopenia-ACLF model. In the training cohort， the sarcopenia-ACLF model had a larger AUC than MELD-Na score in predicting 90-day mortality in patients with ACLF （0.80 vs 0.73， Z=1.97， P=0.049）. In the test cohort， the sarcopenia-ACLF model had a significantly larger AUC than MELD score （0.79 vs 0.69， Z=2.70， P=0.007） and MELD-Na score （0.79 vs 0.68， Z=2.92， P=0.004）. The calibration curve showed that the model had good calibration ability， with a relatively good consistency between the predicted risk of mortality and the observed results. The DCA results showed that within a reasonable range of threshold probabilities， the sarcopenia-ACLF model showed a greater net benefit than MELD and MELD-Na scores in both the training cohort and the test cohort. ConclusionThe sarcopenia-ACLF model developed in this study provides a more accurate tool for predicting the risk of 90-day mortality in ACLF patients， which provides support for clinical decision-making and helps to optimize treatment strategies.

This study aims to explore the mechanism of lung and gut protection by Tanreqing Capsules on the mice infected with influenza virus based on "the lung-gut axis". A total of 110 C57BL/6J mice were randomized into control group, model group, oseltamivir group, and low-and high-dose Tanreqing Capsules groups. Ten mice in each group underwent body weight protection experiments, and the remaining 12 mice underwent experiments for mechanism exploration. Mice were infected with influenza virus A/Puerto Rico/08/1934(PR8) via nasal inhalation for the modeling. The lung tissue was collected on day 3 after gavage, and the lung tissue, colon tissue, and feces were collected on day 7 after gavage for subsequent testing. The results showed that Tanreqing Capsules alleviated the body weight reduction and increased the survival rate caused by PR8 infection. Compared with model group, Tanreqing Capsules can alleviate the lung injury by reducing the lung index, alleviating inflammation and edema in the lung tissue, down-regulating viral gene expression at the late stage of infection, reducing the percentage of neutrophils, and increasing the percentage of T cells. Tanreqing Capsules relieved the gut injury by restoring the colon length, increasing intestinal lumen mucin secretion, alleviating intestinal inflammation, and reducing goblet cell destruction. The gut microbiota analysis showed that Tanreqing Capsules increased species diversity compared with model group. At the phylum level, Tanreqing Capsules significantly increased the abundance of Firmicutes and Actinobacteria, while reducing the abundance of Bacteroidota and Proteobacteria to maintain gut microbiota balance. At the genus level, Tanreqing Capsules significantly increased the abundance of unclassified＿f＿Lachnospiraceae while reducing the abundance of Bacteroides, Eubacterium, and Phocaeicola to maintain gut microbiota balance. In conclusion, Tanreqing Capsules can alleviate mouse lung and gut injury caused by influenza virus infection and restore the balance of gut microbiota. Treating influenza from the lung and gut can provide new ideas for clinical practice.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Lung/metabolism* ; Mice, Inbred C57BL ; Capsules ; Orthomyxoviridae Infections/virology* ; Gastrointestinal Microbiome/drug effects* ; Male ; Humans ; Female ; Influenza A virus/physiology* ; Influenza, Human/virology*

OBJECTIVE: To analyze the effectiveness of three internal fixation methods, namely hollow screw combined with Kirschner wire tension band, hollow screw combined with anchor nail, and modified 1/3 tubular steel plate, in the treatment of avulsion fracture of tibial tubercle (AFTT) in adolescents. METHODS: Between January 2018 and September 2023, 19 adolescent AFTT patients who met the selection criteria were admitted. According to different internal fixation methods, patients were divided into group A (8 cases, hollow screw combined with Kirschner wire tension band), group B (6 cases, hollow screw combined with anchor nail), and group C (5 cases, modified 1/3 tubular steel plate). There was no significant difference in the baseline data of age, gender, side, cause of injury, Ogden classification, and time from injury to operation among the three groups ( P>0.05). The range of motion (ROM), weight-bearing time, normal activity time of knee joint, and the hospital for special surgery (HSS) score at last follow-up were recorded and compared among the three groups. Recorded whether the fracture was displaced, whether the fracture line was blurred at 1 month after operation, whether there was epiphyseal dysplasia, and whether there was incision infection and other complications. RESULTS: There was no significant difference in hospital stay between the groups ( P>0.05). All patients were followed up 10-24 months, with an average of 14.3 months; there was no significant difference between the groups ( P>0.05). All the incisions healed well without soft tissue irritation or fracture nonunion, and no limb shortening deformity or epiphyseal dysplasia was found during follow-up. At 1 month after operation, the knee joint ROM and hospitalization expenses in group A were better than those in groups B and C, the fracture healing time, knee joint weight-bearing time, and normal activity time of knee joint were better than those in group C, and the hospitalization expenses in group C were better than those in group B, with significant differences ( P<0.05); there was no significant difference in the other indicators between the groups ( P>0.05). In group A, the fracture line was blurred 1 month postoperatively, the fracture ends were in close contact, and there was no fracture displacement; in groups B and C, the fracture line was clear in 2 cases, and 1 case in group C had slight fracture displacement; except for 1 case in group B, there was no fracture split in the other two groups. There was no significant difference in the incidences of blur of fracture line, fracture displacement, and intraoperative bone split between the groups at 1 month after operation ( P>0.05). At last follow-up, the HSS scores of knee joints in the three groups were excellent and good, and there was no significant difference between the groups ( P>0.05). CONCLUSION Hollow screw combined with Kirschner wire tension band technique is effective in treating adolescent AFTT, which has the advantages of stabilizing fracture, accelerating fracture healing and rehabilitation, early feasible knee joint functional exercise, and reducing hospitalization expenses.
Humans ; Tibial Fractures/surgery* ; Adolescent ; Male ; Fracture Fixation, Internal/instrumentation* ; Female ; Fractures, Avulsion/surgery* ; Bone Wires ; Bone Screws ; Bone Plates ; Treatment Outcome ; Range of Motion, Articular ; Bone Nails ; Weight-Bearing ; Fracture Healing

OBJECTIVE: To explore effectiveness of intermediate screw assisted reduction and fixation technique in the treatment of Schatzker type Ⅴ and Ⅵ tibial plateau fractures. METHODS: A clinical data of 22 patients with Schatzker type Ⅴ and Ⅵ tibial plateau fractures, who were admitted between June 2022 and December 2023 and met the selection criteria, was retrospectively analyzed. During operation, the intermediate screw assisted reduction and fixation technique was used. There were 14 males and 8 females with an average age of 46.3 years (range, 17-65 years). The fractures were caused by traffic accident in 13 cases, by falls in 5 cases, and by falling from height in 4 cases. According to Schatzker classification criteria, 9 cases were type Ⅴ and 13 cases were type Ⅵ. The interval between injury and operation was 5-12 days (mean, 7.9 days). The operation time, intraoperative fluoroscopy times, and length of hospital stay were recorded. The range of motion of knee joint and Hospital for Special Surgery (HSS) score were recorded at last follow-up. X-ray films were taken to review the fracture healing. Rasmussen score, tibial plateau varus angle (TPVA), and posterior tibial slope (PTS) were estimated before operation, at immediate after operation, and at last follow-up in order to evaluate the fracture reduction effect and postoperative outcome. RESULTS: The operation time was 85-140 minutes (mean, 103.9 minutes). Intraoperative fluoroscopy was performed 7-15 times (mean, 10.1 times). All incisions healed by first intention after operation, and no complication such as nerve or blood vessel injury occurred. The length of hospital stay ranged from 8 to 17 days (mean, 12.4 days). All patients were followed up 10-22 months (mean, 14.8 months). At last follow-up, the range of motion of knee joint was 110°-140° (mean, 125°). HSS score was rated as excellent in 16 cases, good in 5 cases, and fair in 1 case, with an excellent and good rate of 95.5%. X-ray films reexamination showed that all fractures healed with the healing time of 12-17 weeks (mean, 14.8 weeks). No internal fixation failure occurred. Rasmussen score, TPVA, and PTS at immediate after operation and at last follow-up were significantly superior to those before operation ( P<0.05). And there was no significant difference between immediately after operation and last follow-up ( P>0.05). CONCLUSION The treatment of Schatzker type Ⅴ and Ⅵ tibial plateau fractures with intermediate screw assisted reduction and fixation technique is reliable, which can reduce the difficulty of reduction and fixation, improve the efficiency of reduction and fixation, reduce the operation time, achieve satisfactory reduction and fixation effect and postoperative prognosis, and achieve good recovery of knee joint function.
Humans ; Tibial Fractures/diagnostic imaging* ; Male ; Female ; Adult ; Middle Aged ; Bone Screws ; Fracture Fixation, Internal/instrumentation* ; Retrospective Studies ; Adolescent ; Aged ; Young Adult ; Treatment Outcome ; Range of Motion, Articular ; Operative Time ; Length of Stay ; Tibial Plateau Fractures

OBJECTIVE: To evaluate the effectiveness of V-shaped stealth decompression technique using ultrasonic bone scalpel in anterior surgery for adjacent two-level cervical spondylosis. METHODS: A clinical data of 41 patients with adjacent two-level cervical spondylosis, who admitted between January 2020 and December 2023 and met the selection criteria, was analyzed retrospectively. Among them, 22 cases were treated with anterior cervical discectomy and fusion (ACDF) assisted by V-shaped stealth decompression technique using ultrasonic bone scalpel (group A) and 19 cases with anterior cervical corpectomy and fusion (ACCF) (group B). There was no significant difference between the two groups in age, gender, disease duration, surgical segment, preoperative Japanese Orthopedic Association (JOA) score, neck dysfunction index (NDI), pain visual analogue scale (VAS) score, and the anteroposterior diameter of the spinal canal in the responsibility space of axial CT ( P>0.05). The operation time, intraoperative blood loss, postoperative drainage volume, hospital stay, complications during follow-up, JOA score, NDI, and VAS score at last follow-up, and the incidences of intervertebral fusion at 3 months after operation, and cage subsidence at last follow-up were compared between the two groups. RESULTS: The operations in the two groups were successfully completed. The operation time, intraoperative blood loss, postoperative drainage volume, and hospital stay in group A were significantly less than those in group B ( P<0.05). Two cases (9.1%) in group A and 4 cases (21.1%) in group B developed complications, with no significant difference in the incidence between the two groups ( P>0.05). All patients in the two groups were followed up 6-12 months (mean, 9.3 months). There was no significant difference in follow-up time between the two groups ( P>0.05). At last follow-up, the JOA score and VAS score in both groups significantly improved when compared with those before operation ( P<0.05). The change values of VAS score and the improvement rate of JOA score in group A were significantly superior to group B ( P<0.05). There was no significant difference in the change values of NDI and JOA score between the two group ( P>0.05). Imaging reexamination showed that the rate of intervertebral fusion at 3 months after operation was significantly higher in group A (81.8%) than in group B (52.6%) ( P<0.05), and all patients obtained bony intervertebral fusion at last follow-up. At last follow-up, 2 cases (9.1%) in group A and 11 cases (57.9%) in group B had cage sinking, and the difference in the incidence was significant ( P<0.05). No loosening or fracture of internal fixators occurred in all patients. CONCLUSION Using ultrasonic bone scalpel can transform single vertebral ACCF into two-segment ACDF in anterior cervical spondylosis surgery. The V-shaped stealth decompression technique is safe and efficient, with the advantages of minimal trauma, fewer postoperative complications, and rapid recovery of patients.
Humans ; Spondylosis/surgery* ; Male ; Female ; Middle Aged ; Decompression, Surgical/instrumentation* ; Retrospective Studies ; Cervical Vertebrae/surgery* ; Spinal Fusion/instrumentation* ; Diskectomy/instrumentation* ; Adult ; Treatment Outcome ; Aged ; Ultrasonic Surgical Procedures/methods* ; Operative Time

Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*