Hongting Jin

The efficacy of targeted intraarterial delivery of concentrated autologous bone marrow containing mononuclear cells in the treatment of osteonecrosis of the femoral head: a five year follow-up study.

OBJECTIVE To investigate the efficacy and safety of targeted delivery of autologous bone marrow mononuclear cells (BMMCs), which are highly enriched with mesenchymal stem cells (BMMSCs), via medial circumflex femoral artery in the treatment of osteonecrosis of the femoral head (ONFH). METHODS 62 patients (78 hips) with ONFH were recruited in this study. All of these patients were treated with BMMCs perfusion via medial circumflex femoral artery. The concentrated BMMCs (30-60ml) were gained from autologous bone marrow (100-200ml) harvested from anterior iliac crest and then were intra-arterially perfused into the femoral head. Ficat stage was used to classify the radiological stage of ONFH. Harris hip score was used to evaluate the clinical symptoms of osteonecrosis. Ficat stage and Harris hip scores were assessed at onset of treatment at 6, 12, 24, 36, 48 and 60months after the initial treatment. Total hip arthroplasty (THA) was also assessed as an endpoint at each follow-up. RESULTS A follow-up on the patient was done at the end of five years, and 92.31% (72 of 78) of hips achieved a satisfactory clinical result while only 6 hips (7.69%) progressed to clinical failure and required THA. Radiological progression was noted in 34 of 78 hips (43.59%); the overall rate of collapse was 38.24% (26 of 68 hips) in stage-I and stage-II hip combinations and 12.5% (2 of 16) in stage-I hips and 46.15% (24 of 52) in stage-II hips. The mean time of conversion to THA was 3years (1 to 5years) and the average time to collapse were 3.5years (1-5years). The mean Harris hip score increased from 59 points at baseline to 75 points at 12months, 82 points at 24months, 81 points at 36months, 79 points at 48months and 74 points at 60months. Five years after the treatment, 3 of 10 hips (30%) in stage-III had deteriorated to clinical failure whereas only 3 of 68 hips (4.41%) in stage-I and II combination had progressed to clinical failure (p<0.05). Kaplan-Meier survival analysis showed a significant difference in the time to failure between the pre-collapse hips (Ficat stage-I and II) and the post-collapse hips (Ficat stage-III) at five years follow-up (Log-rank test; p<0.01). No complication was found in any patients. CONCLUSIONS Autologous BMMSC perfusion via the medial circumflex femoral artery can relieve symptoms, improve hip function and delay the progression of ONFH. The clinical outcome is better when it is applied prior to the collapse. This work demonstrates that autologous BMMSC perfusion via the medial circumflex femoral artery is a safe, effective and minimally invasive treatment strategy for early-stage ONFH.

Development and assessment of a complete-detoxication strategy for Fuzi (lateral root of Aconitum carmichaeli) and its application in rheumatoid arthritis therapy

ETHNOPHARMACOLOGICAL RELEVANCE Fuzi (lateral root of Aconitum carmichaeli) is a popular traditional Chinese medicine well known for its both therapeutic and high-toxic activities. Its toxic alkaloid ingredients, mainly aconitine, mesaconitine, and hypaconitine, are responsible for the high toxicity. However, to date, no detoxication strategy is available to completely eliminate Fuzis toxicity, and, whether Fuzis efficacy could be kept after detoxication, remain unknown and debatable. MATERIALS AND METHODS The purpose of this study was to establish and validate a complete-detoxication strategy for Fuzi via acute toxicity test, to clarify the detoxication mechanism by HPLC and titrimetric analyses, and to evaluate the therapeutic effect of detoxicated Fuzi on adjuvant arthritis (AA). Three processed Fuzi (Bai-fu-pian) with 30-min, 60-min, and 120-min decoctions, respectively, named dBfp-30, dBfp-60, and dBfp-120, were prepared for this study. For the acute toxicity test, their oral doses to male and female Kunming mice were up to 70-190g/kg body weight, and their toxicological profiles were evaluated by median lethal dose (LD50), maximal tolerance dose (MTD), minimal lethal dose (MLD), no-observed-adverse-effect-level (NOAEL), and time-concentration-mortality (TCM) modeling methods using a 14-day schedule with up to five doses. The HPLC analysis was performed to determine the detoxication-induced changes in composition and amount of aconitine, mesaconitine and hypaconitine in Fuzi, whilst the titrimetric method was adopted to estimate the amount changes of Fuzis total alkaloids. AA model was established by incomplete Freunds adjuvant injection in Wistar rats, and the animals physiological (body weight, food intake, etc.), clinical (hind paw volume), and immunological (IL-1 and TNF-α) parameters were assessed as markers of inflammation and arthritis. RESULTS With increasing decoction time, the acute toxicity of detoxicated Fuzi became decreased in the following order: dBfp-30 (LD50 of 145.1g/kg; MTD of 70g/kg; MLD of 100g/kg; NOAEL of 70g/kg) >dBfp-60 (too large LD50; MTD of 160g/kg; MLD of 190g/kg; NOAEL of 100g/kg) >dBfp-120 (no LD50; unlimited MTD; unlimited MLD; NOAEL of 130g/kg). dBfp-30 and dBfp-60 displayed the toxicity at a dose-dependent manner with maximum mortalities reaching 100% and 50% respectively, whereas no mortality or signs of intoxication was induced by dBfp-120. The chemical analyses revealed a dramatic reduction of the toxic alkaloids as well as total alkaloids in Fuzi after the detoxication, from which no level of aconitine and only minimum residual of mesaconitine (0.56±0.02μg/g) and hypaconitine (8.73±0.13μg/g) were detected in dBfp-120. However, no significant difference of total alkaloid amount was found among dBfp-30, dBfp-60, and dBfp-120 (P>0.05), suggesting an equivalent conversion from toxic alkaloids to its non-toxic derivants in dBfp-120. Further, also no significant differences were seen among dBfp-30, dBfp-60, and dBfp-120 for the therapeutic effects on physiological, clinical, and immunological parameters in AA rat, indicating that dBfp-120 is of non-toxicity and efficacy. CONCLUSIONS A complete-detoxication strategy has been developed successfully for ensuring the safe and effective use of Fuzi. The detoxication mechanism associated with elimination of toxic alkaloids has kept Fuzis efficacy, indicating a non-interdependent relationship between its efficacy and toxicity. This is the first report on such an optimal detoxication strategy and on the application of detoxicated Fuzi in AA. It may provide in depth understanding to the toxicological and pharmacological profiles of Fuzi and further benefit the herbal drug development with safety and efficacy for disease especially RA therapy.

Osteoarthritis Pathogenesis: A Review of Molecular Mechanisms

Osteoarthritis (OA), the most prevalent chronic joint disease, increases in prevalence with age, and affects majority of individuals over the age of 65 and is a leading musculoskeletal cause of impaired mobility in the elderly. Because the precise molecular mechanisms which are involved in the degradation of cartilage matrix and development of OA are poorly understood and there are currently no effective interventions to decelerate the progression of OA or retard the irreversible degradation of cartilage except for total joint replacement surgery. In this paper, the important molecular mechanisms related to OA pathogenesis will be summarized and new insights into potential molecular targets for the prevention and treatment of OA will be provided.

The effects of Liuwei Dihuang on canonical Wnt/β-catenin signaling pathway in osteoporosis.

ETHNOPHARMACOLOGICAL RELEVANCE The Liuwei Dihuang (LWDH), a wellknown classic traditional Chinese medicine formula, consists of six herbs including Rehmannia glutinosa Libosch. (family: Scrophulariaceae), Cornus officinalis Sieb. (family: Cornaceae), Dioscorea opposite Thunb. (family: Dioscoreaceae), Alisma orientale (G. Samuelsson) Juz (family: Alismataceae), Poria cocos (Schw.) Wolf (family: Polyporaceae) and Paeonia suffruticosa Andrews (family: Paeoniaceae). It has been used clinically in the treatment of many types of diseases with signs of deficiency of Yin in the kidneys for more than 1000 years in China. The purpose of this study was to observe the effects of LWDH on canonical Wnt/β-catenin signaling pathway in osteoporosis. MATERIALS AND METHODS Osteoporosis model was induced by ovariectomy (OVX) in 8-week-old female Sprague-Dawley (SD) rats. After 12 weeks of treatment with LWDH by intragastric administration, the rats were put to death in batch. The changes of alkaline phosphatase (ALP), osteocalcin (BGP) and estradiol (E2) in serum were determined, bone mineral density (BMD) and histomorphology of right femur were observed, biomechanics of lumbar vertebra were measured, and the expression of Lrp-5, β-catenin, Runx2, Osx involving the canonical Wnt/β-catenin signaling pathway were detected by RT-PCR. In addition, osteoblasts isolated from neonatal rat calvariae were used in this study to investigate the effects of LWDH on the canonical Wnt/β-catenin signaling pathway. Cell proliferation and differentiation were observed by the MTT test, ALP activity and calcified nodules. The expression of Lrp-5, β-catenin, Runx2, Osx mRNA of cells were also detected. All the data were analyzed by SPSS 13.0. RESULTS Twelve weeks of treatment with LWDH could significantly decrease the level of ALP and BGP in serum, increase the BMD of femurs, and improve the biomechanical capabililty of vertebral body in maximum loading and elastic modulus. Concerning histomorphology, we found ordered arrangement of trabeculae, slightly thinning of trabeculae and none obvious slight fractures in femurs after twelve weeks of treatment with LWDH. In osteoblast, serum containing LWDH elicited significantly increase in cell viability (at day 6), alkaline phosphatase activity (at days 2, 4 and 6) and amount of calcified nodules. The expression of Lrp-5, β-catenin, Runx2 and Osx involved in the canonical Wnt/β-catenin signaling pathway were significantly up-regulated in the presence of LWDH both in vivo and in vitro experiment. CONCLUSIONS Our results suggest that Liuwei Dihuang could alleviate osteoporosis induced by ovariectomy, in part, through up-regulation of canonical Wnt/β-catenin signaling pathway of osteoblast.

Combination treatment of biomechanical support and targeted intra-arterial infusion of peripheral blood stem cells mobilized by granulocyte-colony stimulating factor for the osteonecrosis of the femoral head: a randomized controlled clinical trial.

The objective of this study was to determine the benefits of combination treatment with mechanical support and targeted intra‐arterial infusion of peripheral blood stem cells (PBSCs) mobilized by granulocyte–colony stimulating factor (G‐CSF) via the medial circumflex femoral artery on the progression of osteonecrosis of the femoral head (ONFH). Fifty‐five patients (89 hips) with early and intermediate stage ONFH were recruited and randomly assigned to combination treatment or mechanical support treatment (control group). All hips received mechanical support treatment (porous tantalum rod implantation). Then, hips in the combination treatment group were performed targeted intra‐arterial infusion of PBSCs. At each follow‐up, Harris hip score (HHS) and Association Research Circulation Osseous (ARCO) classification were used to evaluate the symptoms and progression of osteonecrosis. Total hip arthroplasty (THA) was assessed as an endpoint at each follow‐up. At 36 months, 9 of the 41 hips (21.95%) in the control group progressed to clinical failure and underwent THA whereas only 3 of the 48 hips (6.25%) in the combination treatment group required THA (p = 0.031). Kaplan‐Meier survival analysis showed a significant difference in the survival time between the two groups (log‐rank test; p = 0.025). Compared to the control group, combination treatment significantly improved the HHS at 36 months (p = 0.003). At the final follow‐up examination, radiological progression was noted in 13 of 41 hips (31.71%) for the control group, but in only 4 of 48 hips (8.33%) for the combination treatment group (p = 0.005). The overall collapse rates were 15.15% (5/33 hips) and 8.11% (3/37 hips) in the control and combination treatment groups, respectively. Targeted intra‐arterial infusion of PBSCs is capable of enhancing the efficacy of biomechanical support in the treatment of ONFH. This clinical trial confirmed that the combination treatment might be a safe and feasible choice for the treatment of early or intermediate stages of ONFH.

Chondroprotective activity of a detoxicated traditional Chinese medicine (Fuzi) of Aconitum carmichaeli Debx against severe-stage osteoarthritis model induced by mono-iodoacetate.

ETHNOPHARMACOLOGICAL RELEVANCE Fuzi is an effective but toxic traditional Chinese medicine (TCM) derived from Aconitum carmichaeli. In our previous study, detoxicated Fuzi (d-Fuzi) has been originally developed with no toxicity but significant efficacy. However, whether d-Fuzi can be used for therapy of osteoarthritis (OA), remain unknown. MATERIALS AND METHODS Severe OA model was established by intra-articular mono-iodoacetate (MIA) injection (1.25mg) into rats and orally treated with 2g/ml d-Fuzi at a dosage of 7 ml/kg body weight for 28 days. In vivo, the articular radiographic and histopathologic analyses were performed to qualitatively assess the chondroprotective effect of d-Fuzi, followed by quantitative measurements of bone density and Mankin scores. In vitro, such effect on chondrocyte viability after MIA attack was evaluated. Hybrid quadrupole time-of-flight mass spectrometry (QTOF-MS) was performed for chemical analysis of d-Fuzi. RESULTS d-Fuzi was demonstrated to possess chondroprotective activity on MIA-induced OA model by in vivo preventing the articular degeneration and the reducing of bone density and Mankin score, as well as by in vitro promoting the chondrocyte proliferation and inhibiting the MIA-induced chondrocyte damage. A total of 23 compounds were identified in d-Fuzi, most of which were deduced as the non-toxic derivatives of aconite alkaloids. CONCLUSIONS This is the first report regarding chondroprotective effect and chemical profile of d-Fuzi, originally revealing its great anti-OA potential and thereby providing a promising TCM candidate for OA therapy.

Gene Expression Profile of Steroid-induced Necrosis of Femoral Head of Rats

The key to treating steroid-induced necrosis of femoral heads (SINFH) is early diagnosis. Dramatic improvements in diagnosis could be made if the pathogenesis of SINFH was more fully understood; however, the underlying mechanism of this disease is currently unknown. To explore the potential mechanism of SINFH, we performed gene array analysis on a rat model of the disease and compare the expression profile with that of normal rats. A quantitative RT-PCR and immunohistochemistry (IHC) assays were used to confirm the microarray results. Compared to the control group, 190 genes in the experimental group were differentially expressed, with 52 up-regulated and 138 down-regulated. Of these genes, 102 are known (deposited in GenBank), while 88 of them are unknown. The known genes can be divided into several families according to their biological functions, such as oxidative stress, apoptosis, signal transduction, angiogenesis, extracellular matrix, lipid metabolism, and transcription related genes. The results of quantitative RT-PCR and IHC were consistent with gene chip results. Our findings indicate that many genes involved in diverse signaling pathways were differentially expressed between SINFH rats and normal rats. Furthermore, our findings suggest that the development of SINFH is a complicated and dynamic process affected by multiple factors and signaling pathways and regulated by various genes.

The Fate and Distribution of Autologous Bone Marrow Mesenchymal Stem Cells with Intra-Arterial Infusion in Osteonecrosis of the Femoral Head in Dogs

This study aimed to investigate if autologous bone marrow mesenchymal stem cells (MSCs) could treat osteonecrosis of the femoral head (ONFH) and what the fate and distribution of the cells are in dogs. Twelve Beagle dogs were randomly divided into two groups: MSCs group and SHAM operated group. After three weeks, dogs in MSCs group and SHAM operated group were intra-arterially injected with autologous MSCs and 0.9% normal saline, respectively. Eight weeks after treatment, the necrotic volume of the femoral heads was significantly reduced in MSCs group. Moreover, the trabecular bone volume was increased and the empty lacunae rate was decreased in MSCs group. In addition, the BrdU-positive MSCs were unevenly distributed in femoral heads and various vital organs. But no obvious abnormalities were observed. Furthermore, most of BrdU-positive MSCs in necrotic region expressed osteocalcin in MSCs group and a few expressed peroxisome proliferator-activated receptor-γ (PPAR-γ). Taken together, these data indicated that intra-arterially infused MSCs could migrate into the necrotic field of femoral heads and differentiate into osteoblasts, thus improving the necrosis of femoral heads. It suggests that intra-arterial infusion of autologous MSCs might be a feasible and relatively safe method for the treatment of femoral head necrosis.

Synovial sarcoma of the foot: a case report.

A 46-year-old man presented to the authors’ hospitalwith pain and swelling on the medial side of the left foot.When he was 26 years old,a pain had suddenly developedinhisleftfootwithoutanyobviouscause.Thepainalwaysoccurred when he got cold and could be relieved bysoaking in hot water.Five years later,the patient went to alocal clinic as the pain had become more severe and wasoccurring more frequently than before. He was treatedwithIbuprofen,andthepainwasrelieved.Attheageof 43years, he had a surgical excision with a diagnosis of syn-ovialcystbecausehissymptomcouldnotbecontrolledbydrugs.Oneyearlater,becauseof recurrenceof symptoms,he underwent another surgical excision, and pathologicalexamination resulted in a diagnosis of nodular synovitis.However,his symptoms were not eliminated,and affectedhim repeatedly after the second operation. He describedthathisfootgraduallybecamemoreenlargedandswollen,andthatthiswasaccompaniedbyunendurablepainwhileweightbearinginthelastfewmonthsbeforeadmissiontothe authors’ hospital.On admission, physical examination revealed a small,tender, circumscribed, hard mass on the medial side ofthe swollen left foot. The skin was not involved, neitherwas venous dilatation found. The blood supply and sen-sation in the toes of the left foot were normal, whilemovement was impaired. No enlarged local lymph nodeswere found. Radiographs showed bony erosion and peri-osteal reaction in the medial aspect of the second meta-tarsal and the proximal aspect of the first metatarsalbones (Fig. 1). No obvious abnormalities were found onchest radiograph. Incision biopsy and MRI of the leftfoot were performed. A large lobulated soft tissue massmeasuring 7 cm ¥ 5.5 cm ¥ 6 cm was identified. Themass straddled the first to the fourth metatarsal bone(Fig. 2a,b). The histology result was consistent with syn-ovial sarcoma (Fig. 3). Immunochemistry results of epi-thelial membrane antigen (EMA) staining (Fig. 4a) andvideo intensification microscopy (VIM) (Fig. 4b) werepositive and those of CD-34, CD-68, CD-117,Actin, Des,S-100 were all negative, which further supported thediagnosis. The patient underwent left proximal tibialamputation. After the operation, he received adjuvant

Autologous Stem Cells Combined Core Decompression for Treatment of Avascular Necrosis of the Femoral Head: A Systematic Meta-Analysis

Objective This study aims to systematically evaluate the efficacy and safety of core decompression combined transplantation of autologous bone marrow stem cells (CDBMSCs) for treatment of avascular necrosis of the femoral head (ANFH). Methods Randomized controlled trials (RCTs) regarding effectiveness of core decompression combined transplantation of autologous bone marrow stem cells for treating ANFH were searched in 8 comprehensive databases prior to September 2016. The data analysis was performed by using the RevMan version 5.3. Results A total of 11 studies with 507 participants were included. Results showed that CDBMSCs group was more effective than CD group in increasing Harris hip score, decreasing necrotic area of femoral head, collapse of femoral head, and conversion to total hip replacement incidence. In the subgroup analysis, the results did not change in different intervention measure substantially. In addition, the safety of CDBMSCs for ANFH is reliable. Conclusion Based on the systematic review, our findings suggest that core decompression combined transplantation of autologous bone marrow stem cells appeared to be more efficacious in the treatment at early stages of ANFH.