Objective To establish an animal model of osteoporosis combined with liver cirrhosis and conduct preliminary investigation into the effect of liver cirrhosis on bone loss in mice and the underlying mechanisms.Methods The experimental animals were 25 6-week-old female C57BL/6 mice with a body weight of approximately 20-22 g.A comorbidity model of liver cirrhosis and osteoporosis was established in the mice by ovariectomy combined with carbon tetrachloride(CCl4)induction.The mice were randomly assigned to 4 groups(n=5 in each group),including a control group,a liver cirrhosis group,an osteoporosis group,and a cirrhosis and osteoporosis comorbidity group.Pathological changes in the liver were observed via HE staining,Sirius Red staining,and serum liver function indicators.Bone mass and morphological changes were assessed using micro-CT and HE staining.ELISA,Western blot,and immunohistochemistry were performed to assess the expression of insulin-like growth factor-1(IGF-1)in serum and liver tissues.An additional IGF-1 intervention group was established to investigate the potential role of IGF-1 in the comorbidity of liver cirrhosis and osteoporosis,and changes in bone mass and morphology were analyzed via micro-CT and HE staining.Results Compared with the control and osteoporosis groups,the liver cirrhosis and cirrhosis-osteoporosis comorbidity groups exhibited significant inflammatory cell infiltration and collagen fiber deposition in liver tissues,along with markedly increased serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBIL)(P＜0.05).According to the Micro-CT and HE staining results,the cirrhosis-osteoporosis comorbidity group showed reduced bone mass and decreased trabecular numbers in the distal femur compared to those in the osteoporosis group,with the differences being statistically significant(P＜0.05).ELISA,Western blot,and immunohistochemistry demonstrated significantly reduced expression of IGF-1 in the liver and serum of the cirrhosis-osteoporosis comorbidity group(P＜0.05).Notably,exogenous IGF-1 treatment restored bone mass in mice with liver cirrhosis combined with osteoporosis(P＜0.05).Conclusion Through ovariectomy combined with CCl4 induction,a mouse model of liver cirrhosis combined with osteoporosis was successfully established.IGF-1 may serve as a potential molecular mechanism and therapeutic target mediating the liver cirrhosis-osteoporosis comorbidity.

Osteoarthritis(OA),a common age-related chronic disease,is characterized by degenerative changes in the joints and surrounding tissues.Traditionally,research on OA has primarily focused on the pathological changes in articular cartilage and its repair.However,with the advancements in animal disease modeling in recent years,especially the widespread use of spatiotemporally specific transgenic mouse models,scholars have gradually come to realize that the subchondral bone also plays an important role in the occurrence and development of OA.That is,the pathological changes in articular cartilage and bone mutually affect and promote each other,jointly driving the progression of OA,involving such pathological processes as vascular invasion,ectopic calcification,nerve growth,and the occurrence of pain.Given the complexity of cartilage-bone pathological relationship,it is difficult to conduct in-depth research on subchondral bone pathology using clinical human samples,or to simulate the pathological processes of OA through in vitro cell experiments.Therefore,animal models play an irreplaceable role in investigating the pathological mechanisms of OA and developing clinical drugs.This review,in addition to providing an overview of OA animal models,synthesizes the latest progress in animal experimental research on OA,highlighting the active role of the cartilage-bone pathological relationship in OA progression.These new findings provide references for future in-depth investigations and also provide a theoretical basis for developing fundamental strategies for OA prevention and treatment.

Osteoporosis and cardiovascular calcification,two major age-related chronic diseases that China is confronting today,pose serious threats to public health.Previous studies have reported overlapping connections in the pathological processes and molecular mechanisms of these two diseases,particularly concerning inflammation,oxidative stress,and dysregulation of mineral metabolism,and that these two diseases tend to share common pathogenic factors.However,research exploring the comorbidity mechanisms of the two diseases remains limited in both depth and scope,largely due to the lack of widely accepted comorbidity animal models.Herein,we analyzed the latest research findings on the comorbidity mechanisms of vascular calcification and osteoporosis,focusing on summarizing the animal disease models currently in extensive use and the relevant evaluation criteria.We aim to provide new references for comorbidity research models and offer scientific evidence for future studies on pathological mechanisms and the development of new therapeutic strategies.

Objective To investigate the mechanisms underlying regional heterogeneity in the elevating patterns of palatal shelf during mammalian craniofacial development.Methods Using a mouse model of embryonic palatal development,we acquired coronal multi-plane slices of the palatal shelves before elevation(early E13.5),during elevation(late E13.5),and after elevation(early E14.5).Hematoxylin and eosin(HE)staining was performed to compare the morphological changes and spatial correlations between the palate and tongue.Immunofluorescence staining of myosin heavy chain 1(MYH1),a marker found in slow muscle fibers and responsible for muscle contraction and movement,was performed to observe the tongue muscle development characteristics at different stages.We also observed changes in the palatal shelf elevating patterns at early E13.5 in the absence of the tongue through HE-stained in vitro palate organ culture.Further immunofluorescence staining of tenascin-C,an extracellular matrix protein,was performed to evaluate the effect of the tongue on the elevating pattern of the palatal shelf along the anterior-posterior axis.Results HE staining results of the coronal multi-plane slices showed that during the elevation period,from the posterior toward anterior,the coronal height of the tongue decreased,lateral inclination and flattening increased,but the sagittal length of the tongue increased.The elevating pattern of the palatal shelf changed from slow remodeling to rapid flipping,and MYH1 was abundantly expressed in both the internal and external muscle bundles of the tongue during this period.According to findings from in vitro cultivation of palatal organs,the posterior part of the palatal shelf elevated without forming new lateral lingual protrusions in the absence of the tongue.The regional expression pattern of tenascin-C was consistent with that observed before elevation.The posterior palate exhibited an elevation pattern similar to that of the anterior region.Conclusion The tongue may play a crucial role in shaping the posterior morphological remodeling and distinct elevation patterns of the palatal shelf.

Objective:To compare the efficacy of ultrasound combined with endoscopy and ultrasound combined with X-ray guided percutaneous nephrolithotomy(PCNL) in the treatment of complex renal calculi.Methods:The clinical data of 119 patients with complex kidney stones treated by ultrasound combined with endoscopy or ultrasound combined with X-ray guided PCNL in the Second Affiliated Hospital of Anhui Medical University from March 2019 to February 2022 were analyzed retrospectively. According to different guidance methods, they were divided into ultrasound combined with endoscopic guidance group and ultrasound combined with X-ray guidance group.There was no significant difference in age [(53.9±14.2) years vs. (55.6±13.5) years], gender (male/female: 38/21 vs. 30/30), body mass index [(25.0±3.7) kg/m 2 vs. (24.8±3.8)kg/m 2], stone location (left/right: 34/25 vs. 31/29), maximum diameter of renal stones [(31.9±8.3)mm vs. (33.9±13.5)mm], kidney stones maximum cross-sectional area [(601.5±242.5)mm 2 vs. (632.6±278.9)mm 2], number of renal calices involved (3.5±0.9 vs. 3.6±1.3), S. T.O.N.E. scores (9.4±1.0 vs. 9.7±1.4), Guy's grade(Ⅲ/Ⅳ: 45/14 vs. 41/19), preoperative hemoglobin [(125.2±21.5)g/L vs. (125.6±18.4)g/L], serum creatinine[(89.1±33.8) μmol/L vs. (81.9±27.1) μmol/L], urinary tract infection (43/59 vs. 47/60)and positive urinary bacterial culture (12/59 vs.11/60) between the two groups(all P>0.05). The patients in the ultrasound combined with endoscopic guidance group were placed in the modified prone split-leg position. Flexible ureteroscope retrograde into the renal pelvis, combined with ultrasound to determine the best puncture calices. The channels were established and stones were removed under the guidance of ultrasound and endoscopy. In the ultrasound combined with X-ray guidance group, the F5 ureteral catheter was placed retrogradely into the operative side ureter under the lithotomy position. Then the patient changed to prone position and the target calices were punctured under the guidance of ultrasound and X-ray. Through anterograde or retrograde injection of contrast medium, the puncture position was determined to enter in the center of the calicean dome, and the channel establishment process and stone removal are monitored. The operative results and postoperative data were recorded. Results:The average operation time in the ultrasound combined with endoscopic guidance group was significantly shorter than that in the ultrasound combined with X-ray guidance group [(90.2 ± 34.5) min vs. (129.4 ± 43.0) min, P < 0.001]. There was no significant difference in the success rates of initial channel establishment [94.0% (63/67) vs. 87.7% (107/122), P = 0.167], the time of single channel establishment [(7.7 ± 1.9) min vs. (7.7 ± 1.4) min, P =0.765], serum creatinine on the first day after operation[ (89.3±33.6) μmol/L vs. (82.9±27.0) μmol/L, P=0.257] and postoperative hospital stay[(5.3±1.6) d vs.(5.4±1.7) d, P=0.883]. In contrast, patients in ultrasound combined with X-ray guidance group had higher stone free rate [93.3% (56/60) vs. 81.4% (48/59), P=0.049] and lower reoperation rate [3.3% (2/60) vs. 15.3% (9/59), P=0.025]. The mean hemoglobin decrease value of ultrasound combined with endoscopic guidance group was significantly lower than ultrasound combined with X-ray guidance group on the first day after operation [(8.7±6.3) g/L vs. (16.8±6.9) g/L, P<0.001]. The complication rate of ultrasound combined with endoscopic guidance group was significantly lower than that of ultrasound combined with X-ray guidance group [5.1% (3/59) vs. 16.7% (10/60), P = 0.043]. Conclusions:Ultrasound combined with endoscopic guidance PCNL does not need to change body position during operation and has fewer puncture channels, thus saving operation time and reducing complications. It is more suitable for patients with isolated kidney or easy bleeding. Ultrasound combined with X-ray guidance is conducive to the establishment of multi-channel, the stone clearance rate is high and the reoperation rate is low, which is suitable for patients with good health and more renal calices involved with stones.

Microenvironmental biophysical factors play a fundamental role in controlling cell behaviors including cell morphology, proliferation, adhesion and differentiation, and even determining the cell fate. Cells are able to actively sense the surrounding mechanical microenvironment and change their cellular morphology to adapt to it. Although cell morphological changes have been considered to be the first and most important step in the interaction between cells and their mechanical microenvironment, their regulatory network is not completely clear. In the current study, we generated silicon-based elastomer polydimethylsiloxane (PDMS) substrates with stiff (15:1, PDMS elastomer vs. curing agent) and soft (45:1) stiffnesses, which showed the Young's moduli of ~450 kPa and 46 kPa, respectively, and elucidated a new path in cytoskeleton re-organization in chondrocytes in response to changed substrate stiffnesses by characterizing the axis shift from the secreted extracellular protein laminin β1, focal adhesion complex protein FAK to microfilament bundling. We first showed the cellular cytoskeleton changes in chondrocytes by characterizing the cell spreading area and cellular synapses. We then found the changes of secreted extracellular linkage protein, laminin β1, and focal adhesion complex protein, FAK, in chondrocytes in response to different substrate stiffnesses. These two proteins were shown to be directly interacted by Co-IP and colocalization. We next showed that impact of FAK on the cytoskeleton organization by showing the changes of microfilament bundles and found the potential intermediate regulators. Taking together, this modulation axis of laminin β1-FAK-microfilament could enlarge our understanding about the interdependence among mechanosensing, mechanotransduction, and cytoskeleton re-organization.
Cell Adhesion ; Chondrocytes ; Cytoskeleton/metabolism* ; Elastomers/metabolism* ; Laminin/metabolism* ; Mechanotransduction, Cellular

Gap junction (GJ) has been indicated to have an intimate correlation with adhesion junction. However, the direct interaction between them partially remains elusive. In the current study, we aimed to elucidate the role of N-cadherin, one of the core components in adhesion junction, in mediating connexin 43, one of the functional constituents in gap junction, via transforming growth factor-β1(TGF-β1) induction in osteoblasts. We first elucidated the expressions of N-cadherin induced by TGF-β1 and also confirmed the upregulation of Cx43, and the enhancement of functional gap junctional intercellular communication (GJIC) triggered by TGF-β1 in both primary osteoblasts and MC3T3 cell line. Colocalization analysis and Co-IP experimentation showed that N-cadherin interacts with Cx43 at the site of cell-cell contact. Knockdown of N-cadherin by siRNA interference decreased the Cx43 expression and abolished the promoting effect of TGF-β1 on Cx43. Functional GJICs in living primary osteoblasts and MC3T3 cell line were also reduced. TGF-β1-induced increase in N-cadherin and Cx43 was via Smad3 activation, whereas knockdown of Smad3 signaling by using siRNA decreased the expressions of both N-cadherin and Cx43. Overall, these data indicate the direct interactions between N-cadherin and Cx43, and reveal the intervention of adhesion junction in functional gap junction in living osteoblasts.
Cadherins ; Cell Communication ; Connexin 43 ; Osteoblasts ; Transforming Growth Factor beta1

Objective:To discuss the feasibility and clinical efficacy of laparoscopic ureteroplasty with buccal mucosa graft for ureteral stricture.Methods:The clinical data of 10 patients with ureteral stricture admitted to the Second Affiliated Hospital of Anhui Medical University from July 2018 to November 2019 were retrospectively analyzed, including 7 males, 3 females, 5 cases on each side, with an average age of (47.9±7.8)years. All patients had a history of operation related to ureteral calculi. The median value of preoperative serum creatinine was 71(68~610)μmol/L. The status of hydronephrosis and ureteral stricture was evaluated by ultrasonography, CTU and ureteral retrograde angiography.The separation of the renal pelvis on the affected side was (3.1±0.7)cm.There were 7 cases of upper segment stenosis, 2 cases of middle segment stenosis, and 1 case of multiple stenosis. The length of ureteral stenosis was (3.2±0.7)cm. Laparoscopic buccal mucosa ureteroplasty was performed in all 10 cases under general anesthesia. After the stricture ureter segment was separated during the operation, the ureteral stenosis segment was longitudinally cut. According to the stenosis, the buccal mucosa with a length of 3.0-4.5 cm and a width of 1.0-1.5 cm was cut. Buccal mucosa grafts were harvested and placed in the ureter as an anterior onlay with omental wrapping.Results:Ureteral repair was performed laparoscopically in all cases, with no conversion to open and no serious complications. The operative time was (199.2±27.4)min, the blood loss was (101.5±54.7)ml, the median postoperative indwelling time of the drainage tube was 4.5(3.0-7.0)d, and the postoperative hospital stay was (7.9±1.9)days. The patients had clear pronunciation and barrier-free eating one week after the operation.The double J tube was removed one to two months after surgery. The patients were followed up for (11.3±4.2)months after the operation. Follow-up patients underwent imaging and other examinations, which showed significant improvement in hydronephrosis on the affected side, and the median value of renal pelvis separation on the affected side was 1.8(0-2.2)cm. The median value of serum creatinine was 82(66~235)μmol/L. The serum creatinine in 2 patients with renal insufficiency decreased significantly after operation.Conclusions:Laparoscopic buccal mucosa graft ureteroplasty could be a safe and feasible option for the treatment of ureteral stricture with less trauma and rapid recovery. The results of the initial experience are encouraging.

The biophysical properties of the extracellular matrix (ECM) dictate tissue-specific cell behaviour. In the skeleton system, bone shows the potential to adapt its architecture and contexture to environmental rigidity via the bone remodelling process, which involves chondrocytes, osteoblasts, osteoclasts, osteocytes and even peripheral bone marrow-derived stem/stromal cells (BMSCs). In the current study, we generated stiff (~1 014 ± 56) kPa, Young's modulus) and soft (~46 ± 11) kPa silicon-based elastomer polydimethylsiloxane (PDMS) substrates by mixing curing agent into oligomeric base at 1:5 and 1:45 ratios, respectively, and investigated the influence of substrate stiffness on the cell behaviours by characterizing cell spreading area, cell cytoskeleton and cell adhesion capacity. The results showed that the cell spreading areas of chondrocytes, osteoblasts, osteoclasts, osteocytes and BMSCs were all reduced in the soft substrate relative to those in the stiff substrate. F-actin staining confirmed that the cytoskeleton was also changed in the soft group compared to that in the stiff group. Vinculin in focal adhesion plaques was significantly decreased in response to soft substrate compared to stiff substrate. This study establishes the potential correlation between microenvironmental mechanics and the skeletal system, and the results regarding changes in cell spreading area, cytoskeleton and cell adhesion further indicate the important role of biomechanics in the cell-matrix interaction.
Actins ; Cell Adhesion ; Elastic Modulus ; Focal Adhesions ; physiology ; Humans ; Vinculin ; analysis ; metabolism

Objective: To investigate the feasibility and safety of endoscopic combined simultaneous surgery in the modified prone split-leg position for complex renal calculi with ipsilateral ureteral calculi. Methods: The clinical data of 56 cases patients with simultaneous renal and ureteral stones admitted to the Second Affiliated Hospital of Anhui Medical University from January 2016 to March 2019 were retrospectively analyzed. A retrospective analysis was performed on 56 cases of patients with simultaneous renal and ureteral stones who received surgical treatment between January 2016 and March 2019. According to different surgical methods, 56 cases were divided into the modified prone split-leg position group (observation group) and the traditional pre-lithotomy position followed by prone position group (control group). In observation group, the average age of 11 males and 17 females was (54.1±10.2)years. The mean body mass index was (23.8±2.9) kg/m². The location of stones were left in 14 cases and right in 14 cases. The average kidney involvement calyces number was 2.4±0.7.The mean kidney stones maximum cross-sectional area was (870.9±157.7)m². According to the Guy′s classification system, there were 3 cases of grade Ⅰ, 11 case of grade Ⅱ and 14 case of grade Ⅲ in the observation group. The kidney stones S. T.O.N.E scores was 8.7±1.3 and ureteral calculi S. T.O.N.E scores were 13.1± 1.6.In the control group, the average age was (57.0±8.3)years old. The mean body mass index was (24.4±2.9)kg/m². The average kidney involvement calyces number was 2.1±0.7 and the mean kidney stones maximum cross-sectional area was (808.8±189.6)mm². To the kidney stones Guy′s classification, there were 5 cases of grade Ⅰ, 15 case of grade Ⅱ, 7 case of grade Ⅲ and 1case of grade Ⅳ in the control group. The kidney stones S. T.O.N.E scores were 8.5±0.6 and the ureteral calculi S. T.O.N.E scores were 12.4±1.7. The operation time, calculus clearance rate, postoperative hospitalization days, reoperation rate and severity of complications of Clavien-Dindo were statistically compared between the two groups. Results: The study found that the average operation time in the observation group was significantly shorter than that in the control group [(77.8±27.3)min vs.(94.4±22.8)min](P<0.05). In the observation group, 23 patients had complete removal of renal and ureteral calculi, and 3 patients need reoperation. While in the control group, 16 patients had complete removal and 10 patients had reoperation. There were significant differences between the two groups (P<0.05). According to the complication severity of Clavien-Dindo, there were 4 cases of grade Ⅰ and 1 case of grade Ⅱ in the observation group, 4 cases of grade Ⅰ, 2 case of grade Ⅱ and 2 case of grade Ⅲ in the control group. There were no serious complications of grade Ⅳ and Ⅴ in both groups (P>0.05). In observation group, one case accepted the DSA embolization therapy due to the severe bleeding. One case accepted the puncture drainage due to the perinephric effusion. There was no difference in average postoperative hospital stay between the two groups [(6.5±1.2)d vs.(7.0±2.1)d, P>0.05]. Conclusions It is safe and feasible to treat complex renal calculi with ipsilateral ureteral calculi by endoscopic combined simultaneous surgery in the modified prone split-leg position. One position can solve many problems simultaneously, which can significantly reduce the operation time, increase the stone free rates, reduce the reoperation rate and improve the effectiveness of the operation.