This study was aimed to evaluate clinic effect of the layers adjusting external application therapy on knee osteoarthritis (OA). A total of 240 knee OA cases were randomly divided into the treatment group and the control group, with 120 cases in each group. The layers adjusting external application therapy was used in the treatment group. And analgesic plaster of compound of rhizoma arisaematis was used in the control group. The treatment was continued for 21 days. Then, 14-day follow-up was given after the treatment course. There were a total of 35 days. Observations were made on indexes of pain, swelling and joint dysfunction with quantitative scoring. The results showed that the total effective rate was 98.2%in the treatment group, and 83.9%in the control group. The indexes of pain, swelling and joint dysfunction were markedly improved in the treatment group as compared with the control group (P<0.01). It was concluded that the layers adjusting external application therapy was effective on the treat-ment of knee OA.

BACKGROUND:Studies have shown that Weilingxian can maintain and promote the synthesis of proteoglycan and collagen Ⅱ of chondrocyte,and protect artlcular cartilage and postpone the development of osteoarthdtis by inhibiting the level of intedeukin-1(1L-1)possibly.OBJECTIVE:Based on the previous studies,to observe the effect of Weilingxian on the proliferation of rabbit knee articular chondrocyte and transforming growth factor-β1 mRNA expression,and then to explore the role and possible mechanism of Weilingxian in the treatment of osteoarthdtis.METHODS:Knee cartilage was shredded after harvested from New Zealand white rabbits under sterile conditions,and chondrocytes were isolated and cultured by the way Of enzymatic digestion.After identifying by toluidine blue staining,the third-passage calls in the logarithmic growth phase were cultured in vitro and randomly divided into two groups after adherence.The experimental groups were cultured in DMEM with 0.01,0.05,0.1,0.5,and 1.0 mg/mL Weilingxian,while the control group was given with normal medium alone.Chondrecytes morphology was observed under an inverted phase contrast microscope,and the phenotype was identified by toluidine blue staining;Methyl Thiazolyl Tetrazolium(MTT)assay method was adopted to observe the influenca of Weilingxian with difierent concentrations on the proliferation of chondrocytes,and anti-transcription-polymerase chain-type reaction(RT-PCR)was used to assay the expression changes of transforming growth factor-β1 mRNA.RESULTS AND CONCLUSlON:Primary cultured chondrocyte was round-shaped,and most of It adhered after 24 hours,the appearance was polygonal and irregular-shaped;after passage,cell growth was faster than before,the typical appearance was slabstone-like;long spindle-shaped chondrocytes appeared after four generations;after six generations,most cells showed long spindle-shaped fibroblast-like appearance,the rate of growth also slowed down.Extracellular matrix of chondrocytes was stained to be blue by toluidine blue staining,and the nucleus was dark blue.Different concentrations of Weilingxian could promote the proliferation of chondrocytes,effect of 0.5 mg/mL group was significantly,and the peak of proliferation was on the third day.0.05,0.1,0.5,and 1.0 mg/mL Weilingxian group could promote the expression of transforming growth factor-β1 mRNA.and there was no significant difference between four groups(P＞0.05),but the peak was at 0.5 mg/mL group.Weilingxian can promote proliferation of chondrocyte and transfonlling growth factor-β1 mRNA expression,and these may be one of the possible mechanisms that Weilingxian can work in the treatment of osteoarthritis.

AlM: To compare the duration of 0.25% ropivacaine used for sciatic nerve block between type 2 diabetic patients and non-diabetic patients, and to explore the factors affecting the duration of nerve block. METHODS: Sixty eight patients with unilateral lateral malleolus fracture who were to be treated with open reduction and internal fixation were selected from January 2021 to January 2022, aged 20-80 years old, ASA I-III, including 28 diabetic patients and 40 non-diabetic patients. All patients were given 0.25% ropivacaine 20 mL to the superior popliteal sciatic nerve under the guidance of ultrasound. The onset and duration of sensory block were evaluated by blunt needle stimulation. The onset and duration of motor block were evaluated by dorsiflexion and plantar flexion of the operated foot. The interval between the end of the operation and the patientls first request for analgesia was taken as the duration of nerve block analgesia. RESULTS: Compared with non-diabetic patients, the duration of sciatic nerve sensation, motor block and analgesia in diabetic patients were prolonged (P < 0.05). There was no significant difference in the onset time of sciatic nerve sensation and motor block between the two groups (P > 0.05). Linear regression analysis showed that diabetes mellitus, duration of diabetes mellitus, fasting blood glucose and glycosylated hemoglobin were factors affecting the duration of nerve block, and fasting blood glucose was not related to the duration of analgesia. CONCLUSlON: 0.25% ropivacaine can prolong the duration of sciatic nerve block in diabetic patients. The duration of diabetes, diabetes, fasting blood glucose and glycosylated hemoglobin are positively correlated with the duration of block.

OBJECTIVE To investigate the improvement effect and potential mechanism of “Layers adjusting external application” paste on synovial fibrosis （SF） in rats with knee osteoarthritis （KOA）. METHODS Male SD rats were randomly divided into sham operation group， KOA group and Layers adjusting external application group， with 8 rats in each group. KOA model was induced by the anterior cruciate ligament disruption method in KOA group and Layers adjusting external application group. Fourteen days after modeling， the Layers adjusting external application group was given “Layers adjusting external application” paste [Sanse powder （8 g for every 100 cm2）， Compound sanhuang ointment （5 g for every 100 cm2）] on the knee joint， 8 h every day， for 28 d in total. After the last administration， the degree of synovitis and fibrosis in rats was observed， and Krenn scoring was performed in each group. The expressions of collagen Ⅰ， high mobility group protein B1 （HMGB1） and phosphorylated nuclear factor-κB p65 （p-NF-κB p65） were detected in the synovial membrane； the contents of interleukin-1β （IL- 1β）， IL-6 and tumor necrosis factor-α （TNF-α） in serum as well as the expressions of fibrosis-related and HMGB1/Toll-like receptor 4 （TLR4）/NF-κB signaling pathway-related proteins and mRNA were detected in synovial tissue. RESULTS Compared with the sham operation group， the synovial lining cells in the KOA group showed significant proliferation and disordered arrangement， the inflammatory cell infiltration and collagen fiber deposition were obvious； the positive expressing cells of collagen Ⅰ， HMGB1 and p-NF-κB p65 were increased significantly； the contents of IL-1β， IL-6 and TNF-α in serum， the expressions of fibrosis-related protein （transforming growth factor-β， collagen Ⅰ， tissue inhibitor of metalloproteinase 1， α-smooth muscle actin） and their mRNA as well as theexpressions of HMGB1， TLR4 protein and their mRNA， the expressions of p-NF-κB p65 protein and NF-κB p65 mRNA were all increased significantly in synovial tissues of rats （P＜0.01）. Compared with the KOA group， the pathological changes in the synovial tissue of rats in Layers adjusting external application group were significantly improved， and the above quantitative indicators were significantly reversed （P＜0.05 or P＜0.01）. CONCLUSIONS “Layers adjusting external application” paste could significantly improve SF in KOA rats， the mechanism of which may be associated with the inhibition of the activation of HMGB1/ TLR4/NF-κB signaling pathway.