Emodin enhances osteogenesis and inhibits adipogenesis.

BACKGROUND: It has been suggested that the formation of osteoblasts in bone marrow is closely associated with adipogenesis, and the balance between osteogenesis and adipogenesis differentiation of MSCs (mesenchymal stem cells) is disrupted in osteoporosis. In order to improve the treatment of osteoporosis, available agents with roles of regulating the balance is highly desirable. Emodin is a natural anthraquinone derivative extracted from Chinese herbs, which have been used to treat bone diseases for thousands of years. However, the underlying molecular mechanisms of emodin in modulating osteogenesis and adipogenesis remain poorly understood. METHODS: The molecular mechanisms of emodin on the processes of osteogenesis and adipogenesis in ovariectomized mouse and BMSCs (bone marrow mesenchymal stem cells) have been studied. We have analyzed the effects of emodin in vivo and in vitro. Female ICR mice were assigned to three groups: sham group, ovariectomy group, emodin group. Efficacy was evaluated by H&E, immunohistochemical assay and Micro-CT. In vitro, we analyze the effect of emodin--at concentrations between 0.1 µM and 10 µM--on the processes of inducing osteogenesis and inhibiting adipogenesis in BMSCs by ALP, Oil red O staining, real time RT-PCR and western blot. RESULTS: As our experiment shows that emodin could increase the number of osteoblast, BMD (bone mineral density), BV/TV (trabecular bone volume fraction), Tb.N (trabecular number) and Conn.D (connectivity density) of OVX (ovariectomized) mice and decrease the bone marrow fat tissue and adipocytes. The genes and proteins expression of osteogenesis markers, such as Runx2, osterix, collagen type I, osteocalcin, or ALP were up-regulated. While, the genes and proteins involved in adipogenesis, PPARγ, C/EBPα and ap2 were down-regulated. CONCLUSION: It proves that emodin inhibits adipocyte differentiation and enhances osteoblast differentiation from BMSCs.

[Preparation technology optimization of xiaozhong cataplasm by central composite design-response surface method and preliminary clinical effect evaluation].

OBJECTIVE: To optimize the preparation technology of Xiaozhong Cataplasm and to evaluate its preliminary clinical effect. METHODS: The viscosity, residue and appearance were selected as evaluation indexes. The central composite design-response surface methodology was applied to optimize the amounts of Skeleton material, tackifier and crosslinking agent. Clinical effect of Xiaozhong Cataplasm was observed. RESULTS: The optimal formulation was HQ841: PVP k120: PVPP: aluminium glycinate: water: glycerin: ethanol = 4:3:2:0.15: 60:25:10. The effective rate of the cataplasm was 97.22% and 91.67% had marked effect. CONCLUSION: Prepared by the optimal preparation technology, the cataplasm has the moderate viscosity and little residue with good clinical effect.

[Clinical study on core decompression in treating osteonecrosis of the femoral head of the necrotic bone-in different site].

OBJECTIVE: To analyze the clinical effect of decompression and bone grafting on osteonecrosis of the femoral head(ONFH) at different sites of necrotic lesions. METHODS: A total of 105 patients with ARCOⅡstage ONFH admitted from January 2017 to December 2018 were retrospectively analyzed. There were 71 males and 34 females, with an average age of (55.20±10.98) years old. The mean course of all patients was(15.91±9.85) months. According to Japanese Inveatigation Committee (JIC) classification, all patients were divided into 4 types:17 cases of type A, 26 cases of type B, 33 cases of type C1 and 29 cases of type C2. All four groups were treated with decompression of the pulp core and bone grafting. Visual analogue scale(VAS) and Harris hip joint score were used before and at 3, 6, 12, and 24 months after the operation, and the collapse of the femoral head was observed by X-ray examination within 2 years. RESULTS: All 105 patients were successful on operation without complications, and the mean follow-up duration was (24.45±2.75) months. Harris score showed that there was no statistical difference among four groups before surgery and 3, 6 months after surgery (P>0.05);at 12 and 24 months after surgery, there were significant differences among all groups (P<0.01). There were significant differences in intragroup Harris scores at preoperative and postoperative time points among four groups (P<0.01). VAS showed that there was no statistical difference among four groups before and 3, 6 months after surgery (P>0.05);at 12 and 24 months after surgery, there were significant differences among all groups (P<0.01). There were significant differences in VAS at preoperative and postoperative time points among four groups (P<0.01). None of the patients in four groups had femoral head collapse before and 3, 6 months after surgery. At 12 months after operation, there were 3 cases of femoral head collapse in group C and 4 cases in group C2(P>0.05);At 24 months after operation, 1 case of femoral head collapse occurred in group B, 6 cases in group C1 and 8 cases in group C2(P<0.05). CONCLUSION: Core decompression and bone grafting can improve the effect of ONFH and hip preservation. The effect of hip preservation for ONFH is closely related to the location of the osteonecrosis lesion, so the influence of the location of lesion on the effect of hip preservation should be considered in clinical treatment, so as to make better preoperative hip preservation plan.

[Effect of electroacupuncture on the expression of Iba-1 in spinal dorsal horn and hippocampus of chronic pain rats with knee osteoarthritis].

OBJECTIVE: To observe the effect of electroacupuncture(EA) on pain-ralated behaviors, morphology of hippocampus, concentrations of inflammatory cytokines and expression of ionized calcium binding adapter molecule 1(Iba-1) in dorsal horn of the spinal cord and the hippocampus, and brain-derived neurotrophic factor (BDNF) in hippocampus of rats with knee osteoarthritis (KOA), so as to explore the mechanism of EA in improving chronic pain of KOA. METHODS: Forty SD rats were randomly divided into blank group, saline group, model group and EA group, with 10 rats in each group. Monosodium iodoacetate(MIA, 80 mg/mL, 50 µL) was injected into the left knee joint cavity of rats in the model group and EA group to establish the chronic pain model of KOA, while the same volume of normal saline was injected into the left knee joint cavity of rats in the saline group. Rats in the EA group received EA stimulation(2 Hz/100 Hz, 1-2 mA) at left "Yanglingquan"(GB34) and "Neixiyan"(EX-LE4) for 15 min, 14 d after MIA injection. The treatment was given once daily, 5 d as 1 session and 2 sessions of treatment were required. Methanical withdrawl threshold(MWT) and weight-bearing capacity tests on left hind limbs were carried out 1 d before, 7 d，14 d, 20 d and 26 d after MIA injection. At the 27th day, rats were sacrificed and HE staining was used to observe the morphology of hippocampal CA1 area. Concentrations of interleukin(IL)-1ß and tumor necrosis factor(TNF)-α in the left L3-L5 spinal dorsal horn and hippocampal CA1 area were detected by ELISA, the expressions of Iba-1 in the spinal dorsal horn and hippo-campal CA1 area were detected by immunofluorescence, and the expression of BDNF in left hippocampal CA1 area was detected by Western blot. RESULTS: The HE staining results of the hippocampal CA1 area showed reduced number of neurons, unclear cell contour and boundary between nucleus and cytoplasm, and nuclear pyknosis in the model group, which was relatively milder in the EA group. Compared with the blank group, MWT and weight-bearing capacity of rats' left hind limbs, and expression of BDNF protein in hippocampal CA1 area were significantly decreased (P<0.01), while the contents of IL-1ß and TNF-α, the expression of Iba-1 in spinal dorsal horn and hippocampal CA1 area were significantly increased (P<0.01) in the model group. In comparison with the model group, MWT and weight-bearing capacity of rats' left hind limbs, and protein expression of BDNF in hippocampal CA1 area were significantly increased (P<0.01), while the contents of IL-1ß and TNF-α, and the expression of Iba-1 protein in spinal dorsal horn and hippocampal CA1 area were significantly decreased after EA intervention(P<0.01). CONCLUSION: EA at GB34 and EX-LE4 can alleviate the pain-related behaviors of KOA rats. The mechanism might be related to the inhibition of inflammatory reaction mediated by microglia in spinal dorsal horn and hippocampus, and the up-regulation of BDNF expression in hippocampus.

Preventive administration of juanbi capsules for knee osteoarthritis: effects on serum MMP-2 and MMP-9 levels and cartilage repair.

OBJECTIVE: To explore the mechanism of action of Juanbi Capsules, a Chinese medicine for invigorating the kidney and replenishing qi, in preventing osteoarthritis of the knee in rabbits. METHODS: Seventy-two 4-month-old, Japanese long-eared white rabbits were randomly divided into 6 groups: control (group A), model (group B), Chinese drug; high-dose (group C), Chinese drug; mid-dose (group D), Chinese drug; low-dose (group E), and drug control (group F). With the exception of the rabbits in group A, each rabbit was subjected to plaster cast fixation for 6 weeks to induce osteoarthritis. In addition, rabbits were administrated with an intragastric injection of the Chinese drug (groups C, D and E) or an aminoglucose hydrochloride capsule (group F) for 4 weeks. Blood was drawn from the central ear artery for serum MMP-2 and MMP-9 concentrations, and the knee joint cartilage was harvested for gross observation and light microscopy. RESULTS: There were significant differences in serum MMP-2 and MMP-9 concentrations between group B and groups C, D and E (P < 0.05), with no significant differences between groups D and F. Histological results showed various changes in tissue staining with treatment, with osteophyte and bone cyst formation, and superficial erosion in the articular surface of the cartilage; in some cases, the defect reached the mid-layer of the cartilage, and these changes were lower than those in the model group. CONCLUSION: Juanbi Capsules assist in preventing osteoarthritis in the rabbit, possibly by decreasing serum MMP-2 and MMP-9 levels.

Olfactory ensheathing cell transplantation alters the expression of chondroitin sulfate proteoglycans and promotes axonal regeneration after spinal cord injury.

Cell transplantation is a potential treatment for spinal cord injury. Olfactory ensheathing cells (OECs) play an active role in the repair of spinal cord injury as a result of the dual characteristics of astrocytes and Schwann cells. However, the specific mechanisms of repair remain poorly understood. In the present study, a rat model of spinal cord injury was established by transection of T10. OECs were injected into the site, 1 mm from the spinal cord stump. To a certain extent, OEC transplantation restored locomotor function in the hindlimbs of rats with spinal cord injury, but had no effect on the formation or volume of glial scars. In addition, OEC transplantation reduced the immunopositivity of chondroitin sulfate proteoglycans (neural/glial antigen 2 and neurocan) and glial fibrillary acidic protein at the injury site, and increased the immunopositivity of growth-associated protein 43 and neurofilament. These findings suggest that OEC transplantation can regulate the expression of chondroitin sulfate proteoglycans in the spinal cord, inhibit scar formation caused by the excessive proliferation of glial cells, and increase the numbers of regenerated nerve fibers, thus promoting axonal regeneration after spinal cord injury. The study was approved by the Animal Ethics Committee of the Medical College of Xi'an Jiaotong University, China (approval No. 2018-2048) on September 9, 2018.

Effects of preventive administration of juanbi capsules on TNF-alpha, IL-1 and IL-6 contents of joint fluid in the rabbit with knee osteoarthritis.

OBJECTIVE: To probe into the mechanism of the Chinese herbs with functions of reinforcing kidney and supplementing qi for preventing knee osteoarthritis of the rabbit. METHODS: Totally 72 healthy Japan long-ear white rabbits, aged 4 months, were randomly divided into 6 groups, blank group (A), model group (B), high dose Chinese herb group (C), middle dose Chinese herb group (D), small dose Chinese herb group (E), aminoglucose hydrochloride capsule control group (F), 12 rabbits in each group. All the rabbits in the groups, except the group A, were fixed with plaster cast for six weeks to establish rabbit knee osteoarthritis. At the same time of modeling, the different doses of Juanbi Capsules and aminoglucose hydrochloride capsule were administrated intragastrically in the group C, D, E, F, respectively, for 4 weeks, for preventive treatment. In the group B, the rabbit was administrated intragastrically with equal volume of normal saline to the medication groups, twice each day, in the morning and the evening, and in the group A, nothing was administrated. After modeling for 6 weeks, the joint fluid was taken and TNF-alpha, IL-1 and IL-6 contents were detected with ELISA method, and the articular cartilage was taken for macroscopic and microscopic examinations. RESULTS: In all the preventive treatment groups, the articular cartilage color changed to varying degrees with formation of osteophyte and bone cyst, superficial erosion on the chondral articular surface, and the cartilage defect reached to the mid layer in a part of specimens with cartilage exfoliation, but which in the extent were significantly lower than those in the model group. There were significant differences between the group A and B in TNF-alpha, IL-1 and IL-6 contents in the joint fluid (P < 0.05), indicating that the modeling is successful; and there were significant differences as group B compared with the group C,D, E, F, showing that TNF-alpha , IL-1 and IL-6 contents are decreased in all the medication groups; and significant differences between group C, D, E suggests that the increase of Chinese herb doses strengthened the effect of reducing TNF-alpha, IL-1 and IL-6 contents in joint fluid. CONCLUSION: The Juanbi Capsule prevents osteoarthritis possibly through decreasing serum TNF-alpha, IL-1 and IL-6 contents.

Accuracy and Reliability of Computer-aided Anatomical Measurements for Vertebral Body and Disc Based on Computed Tomography Scans.

OBJECTIVE: To assess whether the computed tomography (CT)-based method of three-dimensional (3D) analysis (Mimics) was accurate and reliable for spine surgical anatomical measurements. METHODS: A total of 40 lumbar segments and 32 inter-vertebral discs from eigth adult male cadavers without fractures or deformities fixed with the classical formaldehyde method were included in this research on 5 June 2017. CT scans including seven dimensions: anterior height of the vertebral body (VBHa), middle height of the vertebral body (VBHm), posterior height of the vertebral body (VBHp), width of the upper endplate (EPWu), depth of the upper endplate (EPDu), anterior height of the inter-vertebral disc in the median sagittal plane (IDHa), and posterior height of the inter-vertebral disc in the median sagittal plane (IDHp). They were performed based on uniform conditions (slice thickness: 0.625 mm) using a CT scanner on 8 June 2017. Afterwards, the surgical anatomical measurements were conducted with a Vernier caliper on 12 June 2017. The computer-aided anatomical measurements were conducted by three investigators using Mimics 16.0 to perform 3D reconstructions of CT bone on 16 June 2017. Finally, the length and angle were measured with associated measurement tools, yielding a verified accuracy of 0.01 mm and 0.01°, respectively. Each measurement was repeated three times, and all anatomical data was analyzed using the statistical software and P-value < 0.05 was considered statistically significant. RESULTS: The results showed no statistically significant difference was observed between the surgical anatomical and computer-aided anatomical measurements (P > 0.05) for lumbar vertebra measurements, and the absolute difference between surgical and computer-aided data were all less than 1.0 mm (for the VBHa, VBHm, VBHp, EPWu, and EPDu were 0.12, 0.03, 0.03, 0.31, and 0.03 mm, respectively). Moreover, although the absolute differences of discs was larger than those of lumbar vertebras, no significant differences were detected between the computer-aided and surgical anatomical measurements for the IDHa, as well as IDHp in the vast majority of measurements (P = 0.543, 0.079 or 0.052 for IDHa, and P = 0.212, 0.133 or 0.042 for IDHp). In addition, excellent reliability correlation was observed between the measurements of each investigator, and the reliability coefficients in the intra-groups were all greater than 0.9 except for IDHp (reliability coefficient = 0.892). Additionally, the reliability coefficients were greater than 0.9 for the all between-group correlations, and a significant correlation was also observed. Furthermore, no statistically significant difference for three anatomical values was found in the computer-assisted measurements of the lumbar bone structure (P > 0.05). Similarly, we did not observe a statistical difference in the anatomical data of the lumbar discs from the three measures (P > 0.05). CONCLUSIONS: Computer-aided anatomical measurement for spine based on CT scans presents the high accuracy and reliability for improving spinal surgical procedures.

[Effects of electroacupuncture on pain behavior and pain-related factors in spinal cord dorsal horn and dorsal root ganglia of rats with knee osteoarthritis].

OBJECTIVE: To observe the effect of electroacupuncture(EA) on the pain behavior and prostaglandin E2 (PGE2), calcitonin gene-related peptide (CGRP) and substance P (SP) in the spinal cord dorsal horn and dorsal root ganglia (DRG) of rats with knee osteoarthritis (KOA), so as to explore the mechanisms of EA underlying improvement of chronic pain in KOA rats. METHODS: Thirty-two female SD rats were randomly divided into blank group, control group, model group and EA group, with 8 rats in each group. Rats in the control group were injected with 50 µL of 0.9% sodium chloride solution into the left knee joint cavity, and rats in the model and EA groups were injected with 50 µL of Monosodium iodoacetate in the left knee joint. EA(2 Hz/100 Hz, ＜2 mA) was applied to left "Yanglingquan"(GB34) and "Neixiyan" (EX-LE4) for 15 min, once daily, 5 days a course with a total of 2 courses. Paw withdrawal latency (PWL) and mechanical pain threshold (PWT) were tested by Plantar Test and Von Frey, separately. After the last pain test, the contents of PGE2, CGRP and SP in the left lumbar (L) 3-L5 DRG and L3-L5 spinal dorsal horn were detected by ELISA. RESULTS: Compared with the blank group, the PWL and PWT of the rats in the model group were significantly decreased (P<0.01), and the contents of PGE2, CGRP and SP in the DRG and spinal dorsal horn were significantly increased (P<0.01, P<0.05). There were no statistically significant differences in PWL, PWT, contents of PGE2, CGRP and SP in DRG and spinal dorsal horn between the blank group and the control group (P>0.05). Compared with the model group, the PWL and PWT of rats in the EA group were significantly increased (P<0.01), and the contents of PGE2, CGRP and SP in the DRG and spinal dorsal horn were significantly decreased (P<0.01, P<0.05). CONCLUSION: EA of GB34 and EX-LE4 can reduce the levels of pain-related factors PGE2, CGRP and SP in the DRG and spinal dorsal horn, thereby relieving spinal hyperalgesia in rats with KOA.

Core decompression, lesion clearance and bone graft in combination with Tongluo Shenggu decoction for the treatment of osteonecrosis of the femoral head: A retrospective cohort study.

The aim of this study was to evaluate the clinical effect of core decompression (CD), lesion clearance, and bone graft in combination with Tongluo Shenggu decoction for the treatment of osteonecrosis of the femoral head (ONFH).A total of 75 patients (92 hips), with ONFH at Association Research Circulation Osseous (ARCO) stages II to IIIA, were studied and divided into treatment group and control group. In control group, patients were treated with the CD in combination with autologous or artificial ceramic bone graft. In treatment group, patients were treated with the above method combined with Tongluo Shenggu decoction. Patients were followed-up at 1 month, 6 months, and 24 months after surgery. The visual analogue scale (VAS) scores, Harris Hip Score (HSS), and total effective rates were measured and recorded.The total effective rate of the treatment group was significantly higher than that of the control group (97.2% vs. 89.9%, P < .05). Compared with preoperative, the VAS and HSS scores were both improved at final follow-up, and there was significant difference between 2 groups (P < .01).The combination of CD, lesion clearance, and the bone graft with Tongluo Shenggu decoction is safe and effective for the treatment of ONFH, owing to which it can provide higher postoperative functional outcomes, reduce pain, and achieve smaller osteonecrosis area and better bone changes.

[Effect of Xianlong granules on immunological function in rats of adjuvant arthritis].

OBJECTIVE: To study the effect of Xianlong granules (XLG) on immunological function in the rat of adjuvant arthritis (AA). METHOD: Rats were randomly divided into normal group, AA model group, prednisone group and low, middle and high dose XLG groups, 10 rats in each group. All rats were treated by intragastric administration from the 18 days after arthritis was induced by the complete Freud's adjuvant and the effect of XLG on toes swelling was observed. On the 30th days after modeling, proliferation of the splenic and thymic lymphocytes, and IgG secreted by splenocytes were detected respectively by MTT assay and ELISA. RESULT: Compared with the model group, both the high and middle dose XLG groups had significant therapeutic effects on toes dwelling in the rat of AA (P < 0.05 or P < 0.01); The low, middle and high dose XLG groups strengthened the PHAM-inhibited proliferation of splenic lymphocytes (P < 0.05), and inhibited the PHAM-augmented proliferation of thymic lymphocytes (P < 0.05); XLG did not significantly effect on IgG level secreted by splenocytes in rats of AA. CONCLUSION: XLG can cure toes swelling in rats of AA, which is related with regulation of the abnormal immunlological function.

[Clinical observation on effect of gufusheng capsule in treating early stage Legg-Calve-Perthes disease].

OBJECTIVE: To evaluate the clinical efficacy of Gufusheng capsule (GFSC) in treating early stage Legg-Calve-Perthes disease. METHODS: Adopting randomized single controlled trial, 45 cases with Legg-Calve-Perthes disease in early stage were randomly divided into 2 groups, 23 patients in the control group were treated with arthrectomy hip synovial membrane excision of affected lateral, femoral head decompression and transplantation of muscle-bone flap from ileum, above treatment were also given to the 22 patients in the treated group but combined with orally taking of GFSC. The treatment course for both groups was 6 months. RESULTS: Short-term effect after 6 months' treatment showed that the total effective rate in the treated group and the control group was 90.9% and 78.3% respectively, and the excellent rate was 72.7% and 60.9% respectively. The pain visual analogue scoring, clinical symptom and syndrome scoring markedly decreased before and after treatment, showing significant difference between the two groups (P < 0.05, P < 0.01). Long-term effect after 1-year to 4-year follow-up showed that the total effective rate and excellent rate between the treated group and the control group was significantly different ( chi2 = 8. 5976, P < 0.05). CONCLUSION: GFSC has marked effect in alleviating pain and ameliorating function of hip joint, being an effective compound recipe for treatment of early stage Legg-Calve-Perthes disease. Therefore, it is worthy of researching and developing furthermore.

[Effect of the mixed liquor of danshen and magnesium sulfate injection on IgG and IgM in serum of rabbits with lumbar intervertebral disc herniation].

OBJECTIVE: To explore the effect of the mixed liquor of danshen and magnesium sulfate injection on inflammatory reaction caused by autoimmune response of rabbits with lumbar intervertebral disc herniation. METHODS: Sixty rabbits were divided into blank group (10 rabbits), sham operation group (10 rabbits), model group (40 rabbits) according to method of random digits table. Then model group was divided into made group (group A, 10 rabbits), normal saline group (group B, 10 rabbits), aescin natrium group (group C, 10 rabbits) and danshen and magnesium sulfate injection group (group D,10 rabbits). After model success of lumbar intervertebral disc herniation, different drugs were given to rabbits with lumbar intervertebral disc herniation by ear margin vein. The rabbits of the group C,B,D were respectively given aescin natriu (0.5 mg/kg), normal saline (5 ml/kg), danshen and magnesium sulfate injection (2.0 mg/kg). The rabbits of blank group, sham operation group and group A were not given any disposal. The IgG and IgM level in serum of different groups were determined with ELISA method in fourteen day after model success and drugs given. RESULTS: The level of IgG and IgM in the group A was higher than that of blank group and sham operation group (P < 0.05). The level of IgG and IgM in the group D was lower than that of group B,C (P < 0.05). CONCLUSION: The mixed liquor of danshen () and magnesium sulfate injection could inhibit inflammatory reaction caused by autoimmune response of lumbar intervertebral disc herniation,which provides a new alternative for lumbar intervertebral disc herniation.