Protective effects of Wuwei Xiaodu Drink against chronic osteomyelitis through Foxp3+CD25+CD4+ Treg cells via the IL-2/STAT5 signaling pathway.

To explore the effectiveness and safety of a Chinese medicinal decoction Wuwei Xiaodu Drink (WWXDD) in inhibiting chronic osteomyelitis via regulatory T cells signaling. The effective constitutes of WWXDD and osteomyelitis related genes were screened. Target proteins were cross-validated using the Venny database. GO function and KEGG pathway analysis were performed for target proteins, while pharmacological network was constructed. The bone properties were analyzed by HE staining and the concentrations of immune factors were measured by ELISA. The expression of CTLA-4 and Foxp3 mRNA and STAT5, p-STAT5, CTLA-4 and Foxp3 protein were detected using Real-time PCR and Western blot, respectively. FACS was used to analyze the percentages of cells. A total of 117 genes overlapped between 785 target genes of the active compounds of WWXDD and 912 osteomyelitis related genes. Inflammation-related genes, including IL-6, TNFα, IL-1ß and IL-2 showed high connection degree in the drug-compound-disease-target network. GO function and KEGG pathway analysis revealed that 117 intersection genes mainly enriched in virus infection related pathways, immune related pathways and chemokine signaling pathway. Furthermore, the development of chronic osteomyelitis was suppressed in model rats after treatment with WWXDD. Meanwhile, the concentrations of IL-2 and CD4+CD25+Foxp3 Treg percentages together with the levels of p-STAT5, CTLA-4 and Foxp3 were also down-regulated. Furthermore, IL-2 and WWXDD drug-containing serum exhibited opposite effects on regulating IL-2, IL-10, TGF-ß1, Foxp3, CTLA4 and STAT5. In addition, a STAT5 phosphorylation inhibitor suppressed the expression of Foxp3 and CTLA-4. WWXDD can treat chronic osteomyelitis through suppressing the main regulating factors of Tregs and interfere its immunodepression. Our results bring a new solution for chronic osteomyelitis.

[One-stage compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone for the treatment of chronic calcaneal osteomyelitis].

OBJECTIVE: To explore the treatment of chronic calcaneal osteomyelitis with bone defect after debridement and evaluate its clinical outcomes. METHODS: From June 2009 to June 2011, 52 patients with chronic calcaneal osteomyelitis were treated with stage-one compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone,including 12 females and 40 males with an average age of 43 years old ranging from 18 to 67. According to Cierny-Mader classification, there were 34 cases with stage III and 18 with stage IV. There were 32 cases on right side and 20 on left,with a course of 6 months to 3 years. The area of soft tissue wound ranged from 3.0 cm x l.5 cm to 23.0 cm x l2.0 cm. The clinical effects were evaluated according to infection controlling, calcium sulfate absorption,bone defect repair and heel functional recovery. RESULTS: All patients were followed up for 2 to 3.8 years (averaged 2.8 years). Primary healing was achieved in 52 patients. Two cases of recurrence were found post-operatively, 1 case in 3 months and another in 5 months,which were cured after a second operation. Bone repair healing was gained in 1.5 to 3.5 months (averaged 2.5 months). Complete radiological absorption of calcium sulfate was found in 1.2 to 3 months(averaged 2.2 months). Local exudation after removal of drainage tube had been persisting in 10 patients for 2 to 3 months, which was consistent with the time when cacium sulfate were totally absorbed. Flap had partial necrosis in 4 cases,and the wounds were closed after appropriate treatment finally. The mean Maryland score was 88.15±7.70. There were excellent results in 32 cases, good in 14, fair in 6. CONCLUSION: A satisfactory short-term clinical results can be gained by one-stage compound grafting of antibiotic-impregnated calcium sulfate and autogenous cancellous bone in chronic calcaneal osteomyelitis, but the long-term results need further follow-up. And much more study is also demanded to reduce the exudation of calcium sulfate.

[Application of vancomycin-loaded calcium sulphate in treatment of osteomyelitis].

OBJECTIVE: To investigate the role of vancomycin-loaded calcium sulphate in the treatment of osteomyelitis. METHODS: We implanted vancomycin-loaded calcium sulphate into 24 patients with traumatic osteomyelitis who were treated in our hospital from February 2008 to February 2010,and then the antibiotic concentrations in the lesions were measured.Bacterial culture results,inflammatory markers,as well as wound healing were observed.X-ray was performed in the location where the vancomycin-loaded calcium sulphate was implanted.The blood vancomycin level as well as liver and kidney functions were determined. RESULTS: The vancomycin concentration in the lesion exceeded the effective therapeutic concentrations and the minimum inhibitory concentration,while the blood concentration was low.The liver and renal function remained normal.The safety profile was good,and the cure rate of osteomyelitis reached 100%. CONCLUSIONS: The vancomycin-loaded calcium sulphate can release high-concentration vancomycin in the diseased sites without causing high blood concentration.Also,it can guide the regeneration of bones.Therefore,it is effective and safe in treating osteomyelitis.

[Small needle-knife for the treatment of heel pain according to its classification].

OBJECTIVE: To treat heel pain with small needle-knife according to its classification,so as to improve the therapeutic effects. METHODS: From August 2005 to December 2008, 200 patients with 266 heel pain were treated according to the author's classification standards. There were 83 males and 117 females, ranging in age from 26 to 72 years, with a mean age of 46 years; the course of the disease ranged from 3 to 36 months, averaged 6.8 months. According to the classification:67 heels with plantar fasciitis type, 61 heels with calcaneal bursitis type, 36 heels with plantar fat pad inflammation-based type, 6 heels with calcaneus high pressure-type, 21 heels with nerve compression type, 75 heels with mixed type. All the patients were treated with small needle-knife by using different methods according to its classification. RESULTS: After 1 month treatment, among the patients with plantar fasciitis-type, 31 heels got an excellent result,36 good;among the patients with calcaneal bursitis type, 32 heels got an excellent result, 29 good; among the patients with plantar fat pad inflammation-based type, 9 heels got an excellent result, 20 good, 4 poor and 3 bad; among the patients with calcaneus high pressure-type, 1 patient got an excellent result, 3 good and 2 poor; among the patients with nerve compression type, 11 heels got an excellent result, 8 good, 1 poor and 1 bad; among the patients with mixed type, 16 heels got an excellent result, 46 good, 5 poor and 8 bad. At the 6th month after treatment, among the patients with plantar fasciitis-type, 21 heels got an excellent result, 40 good,5 poor and 1 bad; among the patients with calcaneal bursitis type, 30 heels got an excellent result, 28 good and 3 poor; among the patients with plantar fat pad inflammation-based type, 15 heels got an excellent result, 18 good, 2 poor and 1 bad; among the patients with calcaneus high pressure-type, 0 patient got an excellent result, 3 good and 3 poor;among the patients with nerve compression type,7 heels got an excellent result, 11 good, 1 poor and 2 bad; among the patients with mixed type, 10 heels got an excellent result, 45 good, 11 poor and 9 bad. CONCLUSION: The causes of heel pain can't be explained by using single pathogenesy,and also can't be treated with one method. The patients with calcaneal pain should be treated with different methods according to classification of heel pain, and thus the therapeutic effects can be improved.