BACKGROUND:Emerging evidence demonstrates that low density lipoprotein receptor-related protein 1 (LRP1)isinvolved in lipid metabolism and regulation of inflammatory reaction.
OBJECTIVE:To explore the effectof lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 on cartilage damage and matrix metaloproteinase 13 in a rat model of osteoarthritis, so as to assess the role of low density lipoprotein receptor-related protein 1 in the pathogenesis of osteoarthritis.
METHODS:Sixty-fourSprague-Dawleyratswere included andramdomlydivided into four groups (n=16 for each): negative control group, no surgery; sham-surgery group, onlythearticular cavity of the knee was exposed; osteoarthritisplus shLRP1 group, rat osteoarthritis models were established by cutting anterior cruciate ligament and removing the medial meniscus partly folowed by an intra-articular injection of lentivirus-mediated siRNA at 2 days after surgery, once a week for 2 consecutive weeks; osteoarthritis group,an intra-articular injection of the negative control lentivirus was performed after surgery. Rats inthe four groups started running on theself-made electric treadmil from 5 days after modeling, 30 minutes per day,totaly 500 meters. Cartilage damage and matrix metaloproteinase 13 expression in cartilage tissues were determined at 2, 4, 6 weeks after surgery.
RESULTS AND CONCLUSION:Gross and pathological observations showed that lentivirus-induced knockdown of lowdensity lipoprotein receptor-related protein 1 aggravatedcartilage damage intherat model of osteoarthritis. At 6 weeks after surgery, Mankin’s scoreand matrix metaloproteinase 13 expression inthecartilage tissues in osteoarthritis plus shLRP1 groupweresignificantly increased compared with other three groups (P< 0.05). These findings indicate that a simulation model of osteoarthritis is developed by cutting anterior cruciate ligament and removing the medial meniscus partly combined with running onthe treadmil. Lentivirus-induced knockdown ofLRP1aggravates cartilage damage in a rat model of osteoarthritis

BACKGROUND:Tumor necrosis factor α, as a pathogenic factor, induces the inflammatory reaction mainlyvia the activation of the nuclear factor kappa B signaling pathway. Low density lipoprotein receptor-related protein 1 (LRP1) is involved in the regulation of the inflammatory reaction induced by cytokines.
OBJECTIVE:To study the effect of knockdown of LRP1 on tumor necrosis factor α-induced inflammatory reaction.
METHODS: Primary cultured rat chondrocytes were transfected with lentivirus-mediated RNA interference to knockdown LRP1 gene. Three days after lentivirus transfection, chondrocytes were pretreated with Bay 11-7082 (10 μmol/L) for 30 minutes prior to the addition of tumor necrosis factor α (30 μg/L) for 30 minutes. Signaling protein and mRNA expressions in chondrocytes were detected by western blot assay and real-time PCR analysis, respectively. Chondrocytes were pretreated with or not Bay 11-7082 (10 μmol/L) 30 minutes prior to the addition of tumor necrosis factor α (30 μg/L) for 12 hours after starvation in DMEM for overnight, and the culture medium was colected for ELISA determination of matrix metaloproteinase 13 level.
RESULTS AND CONCLUSION:Tumor necrosis factor α receptor 1 expression was upregulated in chondrocytes after lentivirus-induced knockdown of LRP1. Increased expression of inducible nitric oxide synthase and activation of the nuclear factor kappa B signaling pathway were found after the addition of tumor necrosis factor α in shLRP1 group. Moreover, increased level of matrix metaloproteinase 13 was determined by ELISA. Taken together, knockdown of LRP1 up-regulates the expression of tumor necrosis factor α-induced inducible nitric oxide synthase and matrix metaloproteinase 13 through the activation of the nuclear factor kappa B signaling pathway.

Clinical data of twenty-six patients with primary tumors involving hip ioint treated surgically in Renmin Hospital of Wuhan University between March 1999 and May 2005 were retrospectively analyzed.The diagnosis of all patients was confirmed by pathohistology.There were 3 cases of chondrosarcomas.6 osteosarcomas,1 synovial sarcoma,14 giant cell tumors and 2 aneurysmal bone cysts.Seventeen cases were treated with custom-made total hip replacement prosthesis,7 with custom-made dipolar femoral head prostheses and 2 with saddle prostheses.They were followed up for 18 months to 6 years with an average of 4 years and 3 monts.Local relapse rate and final limb salvage rate was 40%and 60% in 10 patients with malignant bone tumors:125%and 875%in 16 patients with low-grade malignant bone tumors.According to Enneking(MSTS)evaluation criteria.the average score was 19 with an excellent and good rate of 76.9%.Custom-made artificial hip prosthesis is a satisfactory method in limb salvage operations for patients with malignant or low-grade malignant bone tumors jn hip joint.

Objective To observe the effect of acupuncture on cystathionine gamma-lyase(CSE)and myeloperoxidase (MPO) in the colonic tissues of rats and to explore its mechanism for recovering the function of interstitial cells of Cajal(ICCs)in rats after colonic anastomosis . Methods Thirty SD rats were randomized into normal control group, model group(receiving colonic anastomosis) and acupuncture group. The acupuncture group received foot three-needle therapy on bilateral Zusanli(ST36), Sanyinjiao(SP6) and Taichong(LR3), once a day for 3 days after colonic anastomosis. And then the propulsive rate of the small intestine was measured. The count of ICCs with positively expressive c-kit in rat colonic tissues was measured by immunohistochemistry, the activity of CSE was observed by enzyme-linked immunosorbent assay (ELISA) and the activity of MPO was tested by biochemical method. Results Compared with the normal control group, the small intestinal propulsive rate in the model group was decreased, the number of ICCs with positively expressive c-kit was reduced, while the activities of CSE and MPO were increased(P<0.05). The acupuncture group had higher intestinal propulsive rate, more ICCs with positively expressive c-kit, and lower CSE and MPO activities than the model group(P<0.05).Conclusion Acupuncture can promote the recovering of postoperative gastrointestinal function, and its mechanism may be related to the regulation of CSE and MPO activities in the colonic tissues and to the restoration of ICCs function in the focus with positive c-kit.

OBJECTIVETo investigate the effect and clinical significance of glucocorticoid on CD4+CD25+ regulatory T cells (Tregs) in patients with hepatitis B virus (HBV)-related pre-liver failure.

METHODSThe subjects of this study included 78 patients with pre-liver failure induced by HBV (cases) and 24 healthy individuals (controls). Among the 78 cases, 42 received glucocorticoid treatment and 36 did not. Between-group differences in Tregs (in peripheral blood) were evaluated by flow cytometry and statistical analysis.

RESULTSTwo weeks of glucocorticoid treatment led to an increase in Treg level compared to baseline (before therapy: 2.76 ± 0.73 vs. 3.88 ± 1.60). In addition, after the two weeks of glucocorticoid treatment, the Treg level of improved patients was significantly higher than that measured at baseline (before therapy: 2.70 ± 0.77 vs 3.97 ± 1.59, P < 0.05).

CONCLUSIONGlucocorticoids up-regulate the expression of Treg cells, which may contribute to the immunological mechanism that protects pre-liver failure patients from deterioration of their condition. Careful inspection and monitoring of Treg levels may help improve prognosis of these patients.

Adult ; Case-Control Studies ; Female ; Glucocorticoids ; therapeutic use ; Hepatitis B virus ; Hepatitis B, Chronic ; drug therapy ; immunology ; Humans ; Liver Failure ; immunology ; virology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; cytology ; immunology ; Young Adult

Objective:To investigate the feasibility and clinical efficacy of percutaneous anterior column screwing assisted by blocking screws for pelvic and acetabular fractures.Methods:A retrospective analysis was conducted of the 13 patients who had been admitted from July 2019 to April 2022 for pelvic and acetabular fractures. There were 8 males and 5 females with an age of (49.1±13.3) years, 7 acetabular fractures (6 on one side and 1 on both sides; by the Letournel-Judet classification: 5 anterior column fractures on 6 sides, and 2 transverse and posterior wall fractures on 2 sides), and 6 pelvic fractures (5 complicated with pelvic posterior ring fracture; by the Tile classification: 1 case of type B2, 3 cases of type C1, and 2 cases of type C2). According to the anatomic zones of the anterior column, 5 fractures were at zone Ⅲ, 3 ones at zone Ⅳ, and 6 ones at zone Ⅴ. The time from injury to surgery ranged from 3 to 14 days, averaging (8.2±2.9) days. Anterograde anterior column screwing assisted by blocking screws was performed for all the 13 patients; the posterior ring was fixated with percutaneous sacroiliac joint screws for the 5 patients complicated with pelvic posterior ring fracture. The surgical time, intraoperative fluoroscopy frequency, and intraoperative bleeding volume for insertion of anterior column screws, fracture reduction quality, and hip joint function at the last follow-up were recorded.Results:A total of 14 anterior column screws were inserted percutaneously in the 13 patients. For insertion of anterior column screws, the surgical time was (65.0±10.2) min, the intraoperative fluoroscopy frequency (63.5±14.5) times, and the intraoperative bleeding volume for each screw less than 30 mL. All the incisions healed primarily after surgery, without such complications as iatrogenic neurovascular injury or poor wound healing. All the 13 patients were followed up for (11.1±2.2) months after surgery. In the patient with bilateral acetabular anterior column fractures for which 2 anterior column screws had been inserted, one screw had to be removed due to its displacement at 1 month after surgery; no such complications as loosening of internal fixation or fracture re-displacement was found in the other patients. All fractures healed after (10.2±2.1) months. According to the Matta scoring for quality of fracture reduction, 7 sides were excellent, 5 sides good, and 2 sides poor; according to the Majeed scoring for the 6 patients with pelvic fracture at the last follow-up, the efficacy was rated as excellent in 4 cases and as good in 2 ones; according to the modified Merle d'Aubigné & Postel scoring for the 7 patients with 8 acetabular fractures at the last follow-up, the efficacy was rated as excellent in 4 hips, as good in 3 hips, and as fair in 1 hip.Conclusion:For pelvic and acetabular fractures, minimally invasive percutaneous anterior column screwing assisted by blocking screws can result in fine clinical efficacy, in addition to its easy procedures, safety and reliability.

OBJECTIVE To identify and classify the chemical components and components absorbed into blood of Sishen Pills u-sing ultra-high performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry.METHODS SD rats were divided into blank group and drug administration group.The rats in drug administration group were given water extract of Sishen Pills formula intragastrically,and blank and drug-containing plasma were collected respectively.A Hypersil GOLD VANQUISH column(2.1 mm×100 mm,1.9 μm)was used,with 0.1%formic acid water acetonitrile as the mobile phase,gradient elution,volume flow rate of 0.3 mL·min-1,and column temperature of 35℃.Electrospray ion source(ESI)with positive and negative ion scanning mode was used for chromatographic separation and mass spectrometry data acquisition.The chemical components of Sishen Pills were identi-fied by comparing the exact molecular mass,fragment ion information and relative retention time with the map of reference substance,matching with the self-established database and combining with literature reports.On this basis,the components absorbed into blood of Sishen Pills were analyzed by comparing the blank plasma and drug-containing plasma.RESULTS A total of 181 chemical compo-nents were identified from Sishen Pills,mainly including flavonoids,alkaloids,lignans and other components.A total of 49 prototype blood components were identified from the plasma samples,mainly including flavonoids,alkaloids and other components.CONCLU-SION A variety of chemical components in Sishen Pills and drug-containing plasma are comprehensively,accurately and quickly i-dentified,and all of them are assigned to the various medicinal materials in the prescription.This study provides reference for the qual-ity control,basic research on medicinal effect materials and clinical application of Sishen Pills.

BACKGROUNDGastrodin, as one of the major components extracted from the Chinese herb Gastrodia elata Bl., has many biologic effects, one of which is anti-apoptosis. Apoptosis is considered to be one of the pathogenetic mechanisms in steroid-induced osteonecrosis of the femoral head (ONFH). Therefore, we performed this study to investigate whether gastrodin has the potential to prevent steroid-induced ONFH.

METHODSAll 18 male adult Wistar rats were divided equally into three groups: the steroid group, the gastrodin+steroid group, and the control group. Osteonecrosis was induced by low-dose lipopolysaccharide and subsequent high-dose methylprednisolone. Histomorphometric method was used to determine the incidence of osteonecrosis. Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay was performed to detect apoptotic index of osteocytes and osteoblasts. Real-time PCR and Western blotting were performed to detect mRNA and protein expression of Bax, Bcl-2, and Caspase-3. Fisher's exact probability test and one-way analysis of variance (ANOVA) with Turkey's post hoc test were used to examine significant differences between groups.

RESULTSThe incidence of osteonecrosis in the gastrodin+steroid group (16.7%) was significantly lower than that in the steroid group (83.3%). According to TUNEL assay, the apoptotic indices in the steroid group, the gastrodin+steroid group, and the control group were 91.1%, 27.1%, and 5.4%, respectively, and the differences were significant between groups. Compared with the control group and the gastrodin+steroid group, the mRNA and protein expression levels of Bax and Caspase-3 were significantly higher in the steroid group, but the Bcl-2 mRNA and protein expression levels were significantly lower.

CONCLUSIONGastrodin could prevent steroid-induced ONFH by anti-apoptosis.

Animals ; Apoptosis ; drug effects ; Benzyl Alcohols ; therapeutic use ; Femur Head Necrosis ; drug therapy ; prevention & control ; Glucosides ; therapeutic use ; Lipopolysaccharides ; pharmacology ; Male ; Rats ; Rats, Wistar ; Steroids ; pharmacology

ObjectiveTo investigate the feasibility of ethyl acetate fraction of Ipomoea muricatum （IM-EA） in the prevention and treatment of alcoholic gastric ulcer （GU） and explore its mechanism of action based on network pharmacology and experimental verification. MethodForty SD rats were randomly divided into a control group， a model group， a ranitidine group （2.7 mg·kg^-1）， and low- and high-dose IM-EA groups （30，60 mg·kg^-1） after adaptive feeding for 7 days. The GU model was replicated by hydrochloric acid in absolute ethanol （150 mmol·L^-1） in rats after prophylactic administration for one week. Hematoxylin-eosin （HE） staining and periodic acid-Schiff （PAS） staining were used to preliminarily evaluate the efficacy of IM-EA in the prevention and treatment of GU. Lead compounds of IM-EA were screened out by ADMET， and the SwissTarget platform was used to identify the potential targets for these compounds. GU-related targets were collected through DisGeNET， OMIM， and GeneCards databases， which were mapped to potential IM-EA targets to obtain the potential targets of IM-EA against GU. The STRING database was used to construct the protein-protein interaction （PPI） network to screen the hub targets， and the DAVID platform was used to annotate the biological functions of common targets to explore the underlying mechanism of IM-EA against GU. Autodock Vina software was used for the preliminary verification of the computer simulation. The serum levels of tumor necrosis factor （TNF）-α and interleukin （IL）-6 and the content of prostaglandin E₂ （PGE₂）， matrix metalloproteinase-9 （MMP-9）， and superoxide dismutase （SOD） in the gastric tissues were determined by enzyme-linked immunosorbent assay （ELISA）. The relative expression levels of core proteins in the mitogen-activated protein kinase （MAPK） signaling pathway， such as Jun oncoprotein， extracellular signal-regulated kinase （ERK）， and p38， in the gastric tissues were detected by Western blot. ResultAs revealed by the results of animal experiments， compared with the control group， the model group showed significantly damaged gastric tissues and reduced secretion of gastric mucus. Compared with the model group， the groups with drug intervention showed reduced ulcer areas in the gastric tissues （P<0.01） and improved gastric histopathological status and gastric mucus secretion， suggesting that IM-EA was effective in the prevention and treatment of GU. Sixteen lead compounds of IM-EA were screened out by ADMET， and 257 potential targets of IM-EA against GU were obtained. The hub nodes in the PPI network included targets of TNF-α， protein kinase B1 （Akt1）， tumor protein 53 （TP53）， epidermal growth factor receptor （EGFR）， and ERK. Biological functional annotation and molecular docking results suggested that the MAPK signaling pathway potentially played a key role in the prevention and treatment of GU by IM-EA， which was synergistic with the vascular endothelial growth factor （VEGF） signaling pathway， phosphoinositide 3-kinase （PI3K）/Akt signaling pathway， and nuclear factor （NF）-κB signaling pathway in anti-inflammation， anti-oxidation， and damage repair. The pharmacological experiment results showed that compared with the control group， the model group showed increased serum IL-6 content （P<0.01）， an increasing trend of TNF-α content， increased MMP-9 content in the gastric tissues （P<0.01）， and decreased SOD content （P<0.05）. Compared with the model group， the IM-EA groups showed decreased TNF-α and IL-6 levels in the serum and PGE₂ and MMP-9 levels in the gastric tissues （P<0.01）， and increased SOD content in the gastric tissues （P<0.01）. Compared with the control group， the model group showed up-regulated expression of p-p38， p-Jun， and p-ERK in the gastric tissues （P<0.01） and up-regulated p38 and Jun （P<0.01）. Compared with the model group， the IM-EA groups showed down-regulated p-p38， p-Jun， p-ERK， and p38 in the gastric tissues （P<0.01） and up-regulated relative expression of Jun and ERK （P<0.05）. ConclusionIM-EA has a remarkable effect in the prevention and treatment of alcoholic gastric injury， which may be achieved through the mechanisms of anti-inflammation， anti-oxidation， and wound repair mediated by the MAPK signaling pathway.

ObjectiveTo investigate the mechanism of protective effect of ethanol extracts of Hemsleya chinensis （HC-EE） on hydrochloric acid/ethanol （HCl/EtOH）-induced acute gastric ulcer in rats. MethodLipopolysaccharide （LPS）-induced RAW264.7 cells were used to evaluate inhibitory effect of HC-EE on the production of inflammatory mediators in vitro. A rat acute gastric ulcer model induced by HCl/EtOH （60% ethanol in 150 mmol·L^-1 HCl） was used to evaluate protective effect of HC-EE on acute gastric ulcer. Rats were divided into five groups， including normal group， model group， HC-EE low dose （HC-EE 30， 30 mg·kg^-1） group， HC-EE high dose （HC-EE 60， 60 mg·kg^-1） group and ranitidine （35 mg·kg^-1） group. For model and drug-treated groups， vehicle solvent or drugs were orally administered twice daily for 7 consecutive days before the rats were subjected to HCl/EtOH to induce acute gastric ulcer. After being anesthetized， ulcer surface of each rat was obtained and recorded using electronic imaging technology， and the ulcer inhibition rate was calculated by ImageJ 1.8.0. Hematoxylin-eosin （HE） and periodic acid-Schiff （PAS） staining were used to observe the pathological histological changes in rats. Content of nitric oxide （NO） in cell culture medium was measured by the Griess method. The levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， IL-6， vascular cell adhesion molecule-1 （VCAM-1） and prostaglandin E₂ （PGE₂） in rat serum （or cell culture medium） were determined by enzyme linked immunosorbent assay （ELISA）. The protein expressions of phosphorylation （p）-p38 mitogen activated protein kinase （MAPK）/p38 MAPK and p-nuclear transcription factor-κB （NF-κB） p65/NF-κB p65 in rat gastric tissue were detected by Western blot. ResultIn vitro assay showed HC-EE could significantly down-regulate the expressions of NO， TNF-α， IL-1β， IL-6 and VCAM-1 in LPS-induced cells （P<0.05， P<0.01）. In vivo experimental results showed that， compared with the normal group， gastric tissue of the model group was severely damaged， and the area of gastric ulcer was significantly enlarged， levels of TNF-α， IL-6 were significantly increased （P<0.01）， and the level of PGE₂ was significantly decreased （P<0.01）， the phosphorylation levels of of p38 MAPK， NF-κB p65 in gastric tissue were significantly increased （P<0.01）. Compared with the model group， HC-EE dose-dependently improved HCl/EtOH-induced gastric tissue injury and inflammatory cell infiltration， and it could increase ulcer inhibition rate， significantly decreased the release of TNF-α and IL-6 （P<0.01）， HC-EE 60 group could increase the content of PGE₂ （P<0.05）， and significantly inhibit the phosphorylation levels of p38 MAPK and NF-κB p65 （P<0.05， P<0.01）. ConclusionHC-EE can exert protective effect on HCl/EtOH-induced acute gastric ulcer in rats， and its mechanism may be related to the inhibition of expression of inflammatory mediators mediated by p38 MAPK/NF-κB signaling pathway.