Chinese medicine Di-long (Pheretima vulgaris) and its active fraction exhibit anti-rheumatoid arthritis effects by inhibiting CXCL10/CXCR3 chemotaxis in synovium.

ETHNOPHARMACOLOGICAL RELEVANCE: Di-Long (Pheretima vulgaris) is a classic animal sourced traditional Chinese medicine. It has been used for the treatment of joint inflammation and arthralgia for over two thousand years due to its effects of Tong-Luo-Zhi-Tong (dredging collaterals and alleviating pain). Our previous study showed that Chinese medicine Di-Long has significant anti-rheumatoid arthritis (RA) effects. AIM OF THE STUDY: Considering Di-Long as a potential source of active compounds with specific anti-RA therapeutic effects, this research was to obtain the anti-RA target-specific active fraction from Di-Long extracts (DL), and to further explore the chemical basis and verify the anti-RA mechanism of this active fraction. MATERIALS AND METHODS: Transcriptomic was applied to obtain the main anti-RA targets of DL on human RA fibroblast-like synoviocytes (FLS) and validated by qPCR. The target-corresponding active fraction was isolated from DL by ethanol precipitation and gel chromatography, and analyzed by nanoliter chromatography-mass spectrometry. Anti-RA effects of this active fraction was investigated by collagen-induced arthritis (CIA) in mice, and anti-RA mechanisms were verified in cocultured model of rat FLS and peripheral blood lymphocytes. RESULTS: We confirmed that CXCL10/CXCR3 was the main anti-RA target of DL. The active fraction - A (2182 - 890 Da) was isolated from DL based on its CXCL10 inhibiting effects in RA-FLS. Fraction A contains 195 peptides (192 were newly discovered), 26 of which might be bioactive and were considered to be the chemical basis of its anti-RA effects. Fraction A significantly ameliorated the joint destruction and overall inflammation in CIA mice, and downregulated CXCR3 expression in mice joint. Fraction A inhibited the chemotaxis of Th-cells in rat peripheral blood lymphocytes towards the TNF-α-induced rat FLS through CXCL10/CXCR3 pathway. CONCLUSIONS: Our work indicated that active fraction from DL containing small peptides exhibits promising therapeutic effects for RA through inhibiting CXCL10/CXCR3 chemotaxis.

Combined computational analysis and cytology show limited depth osteogenic effect on bone defects in negative pressure wound therapy.

Background: The treatment of bone defects remains a clinical challenge. The effect of negative pressure wound therapy (NPWT) on osteogenesis in bone defects has been recognized; however, bone marrow fluid dynamics under negative pressure (NP) remain unknown. In this study, we aimed to examine the marrow fluid mechanics within trabeculae by computational fluid dynamics (CFD), and to verify osteogenic gene expression, osteogenic differentiation to investigate the osteogenic depth under NP. Methods: The human femoral head is scanned using micro-CT to segment the volume of interest (VOI) trabeculae. The VOI trabeculae CFD model simulating the bone marrow cavity is developed by combining the Hypermesh and ANSYS software. The effect of trabecular anisotropy is investigated, and bone regeneration effects are simulated under NP scales of -80, -120, -160, and -200 mmHg. The working distance (WD) is proposed to describe the suction depth of the NP. Finally, gene sequence analysis, cytological experiments including bone mesenchymal stem cells (BMSCs) proliferation and osteogenic differentiation are conducted after the BMSCs are cultured under the same NP scale. Results: The pressure, shear stress on trabeculae, and marrow fluid velocity decrease exponentially with an increase in WD. The hydromechanics of fluid at any WD inside the marrow cavity can be theoretically quantified. The NP scale significantly affects the fluid properties, especially those fluid close to the NP source; however, the effect of the NP scale become marginal as WD deepens. Anisotropy of trabecular structure coupled with the anisotropic hydrodynamic behavior of bone marrow; An NP of -120 mmHg demonstrates the majority of bone formation-related genes, as well as the most effective proliferation and osteogenic differentiation of BMSCs compared to the other NP scales. Conclusion: An NP of -120 mmHg may have the optimal activated ability to promote osteogenesis, but the effective WD may be limited to a certain depth. These findings help improve the understanding of fluid mechanisms behind NPWT in treating bone defects.

Transfers of the Ipsilateral C7 Plus the Spinal Accessory Nerve Versus Triple Nerve Transfers for Treatment of C5-C6 Avulsion of the Brachial Plexus.

PURPOSE: To compare the long-term results of transfers of the ipsilateral C7 (IC7) plus spinal accessory nerve (SAN) with those of triple nerve transfers (TNT) using one fascicle of the ulnar nerve to the biceps motor branch (Oberlin's procedure), SAN transferred to the suprascapular nerve, and transfer of the long head of triceps nerve branch to the anterior branch of axillary nerve to treat C5-C6 avulsion of the brachial plexus. METHODS: The IC7 group included 9 patients undergoing transfers of IC7 to the upper trunk and SAN to the suprascapular nerve. Median age at surgery was 26 years and interval between injury and surgery was 2.8 months. Patients were observed for a median of 118 months. The TNT group contained 13 patients, median age 33 years; interval between injury and surgery was 3.1 months. Patients were observed for a median of 103 months. RESULTS: In the IC7 group, median shoulder abduction was 105° and median external rotation of the shoulder was 64°, which was similar to that of the TNT group (89° abduction and 58° external rotation). Eight of nine patients recovered at least M3 (Modified Narakas scale) strength of deltoid in the IC7 group, which was similar to that in the TNT group (11 of 13 patients). Six of nine patients achieved at least Medical Research Council grade 3 (MRC3) strength of biceps in the IC7 group, which was similar to that in the TNT group (11 of 13 patients). Of 4 patients in the IC7 group with a preoperative latissimus dorsi strength of MRC3 or less, 3 gained a deltoid strength of M3 or less, and 3 a biceps strength of MRC2 or less. CONCLUSIONS: Transfers of IC7 plus SAN provide results comparable to those of TNT for treatment of C5-C6 avulsion. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Sensorimotor cortical changes assessed with resting-state fMRI following total brachial plexus root avulsion.

OBJECTIVE: Peripheral nerve injury can induce immediate and long-standing remodelling of the brain cortex, which may affect outcomes of nerve repair. This study examined changes of corresponding cortical representations in patients with brachial plexus injuries. METHODS: Resting-state fMRI was acquired for 13 adult patients with total brachial plexus root avulsion, three of whom underwent second scans 7 or 8 months later. The time of examination ranged from 1 to 16 months after injuries. Nine healthy adults were enrolled as control. Seed-based functional connectivity was performed for all subjects. RESULTS: For nine patients whose first fMRI was performed from 1 to 4 months after brachial plexus injuries, images showed that their cortical maps of sensorimotor areas corresponding to the hand and arm in the hemisphere contralateral to the injured side had much weaker correlation with the supplementary motor area (SMA) than those ipsilateral to the injured side. Symmetrical maps of bilateral cortical sensorimotor areas corresponding to the hand and arm were observed in other four cases with fMRI tested from 7 to 16 months after injuries. For three of the nine patients with asymmetrical cortical representations, second scans indicated symmetric results or even stronger correlation with SMA in the cerebral cortex contralateral to the injured side. CONCLUSIONS: Total brachial plexus root avulsion causes cortical representations of the brachial plexus to undergo a change from an inactive to an active state. This implies that peripheral deafferentation after brachial plexus injuries will induce corresponding cortical representations to be occupied by adjacent non-deafferented cortical territories.

Results of intercostal nerve transfer to the musculocutaneous nerve in brachial plexus birth palsy.

BACKGROUND: Intercostal nerve (ICN) transfer has been one of the main extraplexal nerve transfers in treating brachial plexus root avulsion. This retrospective study evaluated results of ICN transfer for reconstruction of the musculocutaneous nerve (MCN) in brachial plexus birth palsy (BPBP). METHODS: Eighteen boys and 6 girls with BPBP, who had avulsion of at least 2 spinal nerves of the plexus, underwent ICN transfer for reconstruction of MCN, from March 2003 to October 2005. The brachial plexus lesion was diagnosed by clinical assessment, surgical exploration, and intraoperative neurophysiological investigations. The age at surgery ranged from 3 to 11 months of life, with a mean of 5 months. Two intercostals were used for one, 3 intercostals for 9, and 4 intercostals for 14 patients. The intercostals were transferred to MCN in 12 and to the anterior division of the upper trunk in the other 12 cases. RESULTS: Twenty-four children were followed up for 24 to 79 months, with an average of 53 months. No complications were found in the respiratory system. Of 14 transfers with 4 intercostals, biceps gained M4 strength in 8, M3 in 4, and M2 in 2. Of 9 transfers with 3 intercostals, biceps obtained M4 strength in 8 and M3 in 1. One transfer with 2 intercostals got M4 strength of biceps. Twelve patients whose intercostals were transferred to MCN, gained M4 strength of biceps in 11 and M3 in 1, whereas the other 12 patients with intercostals transferred to anterior division of the upper trunk, obtained M4 strength of biceps in 6, M3 in 4, and M2 in 2. The rate of M3 strength or more was 92% and that of M4 was 71%. CONCLUSIONS: ICN transfer is a safe and reliable procedure for reconstruction of the MCN in BPBP. There seems to be no difference of effects between transfers with 3 and those with 4 intercostals. The transferred nerves should be coapted to MCN, rather than a more proximal portion of the plexus. LEVEL OF EVIDENCE: Level III: retrospective comparative study.

Histopathologic study of the neuroma-in-continuity in obstetric brachial plexus palsy.

BACKGROUND: Operative treatment of traction lesions in obstetric brachial plexus palsy is still controversial. The authors analyzed the histopathology of neuroma-in-continuity of the upper trunk by study of the resected neuroma. METHODS: The neuroma-in-continuity of the upper trunk was studied histopathologically in 28 children with Erb palsy who had undergone resection of the neuroma and nerve reconstruction of the plexus at the age of 3 to 11 months. The authors recorded the distribution of myelinated motor nerve fibers and the proportions of collagen and regenerating nerve fibers traveling the neuroma, analyzed the relationship between the percentage of nerve fibers across the neuroma and findings of intraoperative neurophysiologic investigations and the patient's age at surgery, and compared the number of nerve fibers in C5 and C6 proximal to the neuroma with that in their normal counterparts. RESULTS: In the central segment of the neuroma, the structure of the upper trunk was replaced by copious collagen and sporadic nerve fibers wrapped by an undeveloped myelin sheath, and the percentage of collagen was statistically greater than that of the normal upper trunk. The mean percentage of regenerating nerve fibers across the neuroma was 41.83 percent (95 percent confidence interval, 38.69 to 44.69 percent) and this was not statistically correlated with the outcome of intraoperative neurophysiologic investigations or the patient's age at surgery. The number of nerve fibers was statistically less in C5 and C6 proximal to the neuroma than in their normal counterparts. CONCLUSIONS: The nerve structure of the neuroma-in-continuity is substantially damaged in obstetric brachial plexus palsy. Its resection followed by nerve reconstruction of the plexus is favored.

Origination of the brachialis branch of the musculocutaneous nerve: an electrophysiological study.

OBJECTIVE: To test an innovative method to study the origin of a specific nerve or of the nerve fibers innervating a given muscle on the healthy upper limb of a human being and to find the rationale for the brachialis branch of musculocutaneous nerve transfer. METHODS: An intraoperative electrophysiological study was conducted comprising 27 cases of contralateral C7 transfer. The goal of the study was to record compound muscle action potential of the brachialis muscle while various nerve roots of the brachial plexus were stimulated. RESULTS: Analysis of compound muscle action potential suggested that the brachialis branch of the musculocutaneous nerve is composed of fibers from the C5, C6, and C7 nerve roots and that the C5 and C6 nerve roots are the major origin for the brachialis branch of musculocutaneous nerve fibers. CONCLUSION: The technique proposed here was a more direct and functional method of tracing the origin of a specific nerve or of the nerve fibers innervating a given muscle on the healthy upper limb of a live patient.

Contralateral C7 transfer for the treatment of brachial plexus root avulsions in children - a report of 12 cases.

PURPOSE: To retrospectively determine the risks and benefits of contralateral C7 nerve root transfer in infants and children. METHODS: In 12 infants and children with brachial plexus root avulsions from birth injury or other trauma, the common trunk of the contralateral C7 root was transferred to the trunk, division, cord, or nerve branch(es) on the affected side with 2 different types of interposition grafts. The surgery was performed in 1 stage for 5 patients and in 2 stages for 7 patients. RESULTS: Patients were followed up for a mean of 42 months, with a minimum of 21 months. Noteworthy function (> or = M2+, modified British Medical Research Council grading system) was gained in 10 of 12 patients and sensory function (> or = S3, British Medical Research Council grading system) was gained in all patients. Improvements in strength and sensation were accompanied by little synchronous motion and sensibility changes in the donor limb in 7 children, to whom the repaired nerves were those innervating the shoulder and/or elbow or both the musculocutaneous and median nerves. In addition to slight damage to the sensory function of the median nerve, 2 infants also had temporarily reduced shoulder abduction on the healthy side. CONCLUSIONS: For contralateral C7 transfer in infants and children with brachial plexus root avulsions, the deficit created by the procedure is minimal and motor and sensory function is gained. Transfer of the contralateral C7 root to different nerves for a child may improve the quality of functional recovery. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic, Level IV.

[Long-term impact of transfer of phrenic nerve on respiratory system of children: a clinical study of 34 cases].

OBJECTIVE: To study the long-term impact of transfer of phrenic nerve on respiratory system of children. METHODS: Thirty-four children with brachial plexus injury, 25 boys and 9 girls, underwent transfer of phrenic nerve and were divided into 3 groups according to the age when they underwent operation: group of the age of 0 - 12 months (n = 17), group of 13 - 36 months (n = 11), and group of 37 - 60 months (n = 6). Thirty-four sex, height, and body weight-matched healthy children were used as controls. Follow-up, including physical examination, pulmonary function examination (tidal volume, ventilation, etc), blood gas analysis, and chest radiography, was conducted for 4.03 years (3 - 7 years). RESULTS: The values of maximum vital capacity of the group of 0 - 12 months and group of 13 - 36 months were 1.0 L +/- 0.2 L and 1.2 L +/- 0.4 L, both significantly lower than those of the corresponding control groups (1.3 L +/- 0.3 L and 1.4 L +/- 0.5 L, both P < 0.05). The values of one-second vital capacity of the group of 0 - 12 months and group of 13 - 36 months were 0.8 L +/- 0.1 L and 0.9 L +/- 0.1 L, both significantly lower than those of the corresponding control groups (1.0 L +/- 0.1 L and 1.0 L +/- 0.1 L, both P < 0.05). However, the values of the maximum vital capacity and one-second vital capacity of the group of 37 - 60 months were 1.6 L +/- 0.3 L and 1.8 L +/- 0.5 L respectively, both not significantly different from those of the controls (both P > 0.05). The results of blood gas analysis of the 3 operation groups were not significantly different from those of the corresponding controls. Chest radiograph showed that the diaphragm top was raised by 1.93 intercostal spaces (0.5 - 3.5 intercostal spaces) in comparison with the contralateral sides with significant differences between the group of 0 - 12 months and the group of 13 - 36 months and between the group of 0 - 12 months and the group of 37 - 60 months (both P < 0.05). The recurrent respiratory infection rate and of the groups of 0 - 12 months and 13 - 36 months were 47.1% and 27.3% respectively, both significantly higher than that of the group of 37 - 60 months (0%). The thorax deformity rate of the groups of 0 - 12 months and 13 - 36 months were 41.2% and 9.1% respectively, both significantly higher than that of the group of 37 - 60 months (0%). Three of the children in the group of 0 - 12 months (17.6%) had digestive system symptoms. CONCLUSION: Transfer of phrenic nerve operated on children younger than 3 years may cause abnormalities of respiratory system, thorax, and digestive system. The younger the patients the more severe the consequences of the operation. The children older than 3 years tolerate the operation better.

Functional reconstruction of the irreparable upper trunk defect of the brachial plexus--a case report.

We report a successful application of Oberlin's procedure combined with transfer of trapezius and latissimus dorsi with the teres major for reconstruction of elbow flexion as well as abduction and external rotation of the shoulder to a ten-year-old patient, who had a long defect of the left brachial plexus upper trunk caused by resection of the plexiform neuroma.

Selective transfer of the C7 nerve root: an experimental study.

The authors present selective C7 nerve root transfer in a rat model. The musculocutaneous nerve was neurotized by various portions of ipsilateral C7. The latent period and maximum amplitude of evoked motor action potential of the biceps, number of regenerating myelinated nerve fibers, cross-sectional area and wet weight of the biceps, and twitch and tetanic tensions of the biceps were measured at four postoperative intervals. In the early postoperative period (1 and 2 months), nerve regeneration in neurotization with the posterior division or the anterior division of C7 was significantly better than that with the anterolateral fascicles of the anterior division or the phrenic nerve. As the postoperative interval prolonged, the parameters of nerve regeneration in the latter two groups approximated those in the former two groups. This indicated that there were enough regenerating nerve fibers in the anterolateral fascicles of the anterior division and a promising potential for nerve regeneration. The clinical significance of the results lies in the design of selective C7 transfer which, using the anterolateral fascicles of the anterior division, could preserve the function of the muscles innervated by the posterior division to the greatest extent, and provide sufficient donor outflow as well. It is therefore a new option for C7 transfer.

Comparative experimental study on treatment outcome of nerve transfer, using selective C7 nerve root vs. phrenic nerve.

The treatment outcome of nerve transfer using the C7 nerve root or phrenic nerve was compared in a rat experiment. One hundred and twenty SD rats were divided into two groups, one undergoing phrenic nerve transfer to the musculocutaneous nerve, and the other partial ipsilateral C7 (anteriolateral fascicles of the anterior division) to the musculocutaneous nerve. Neurotization outcomes of the two groups were evaluated by comparing the electrophysiologic, histologic, and myophysiologic changes of the biceps muscle. No significant differences were found between parameters from the phrenic nerve transfer group and those from the ipsilateral C7 nerve transfer group. This indicates that the treatment outcome of selective ipsilateral C7 transfer is comparable to that of phrenic nerve transfer. It is the surgery of choice in treating brachial plexus upper-trunk avulsion accompanied by phrenic nerve injury.

Electrophysiologic study of influence on C7 innervated muscles after transection of different C7 fascicles.

An experimental study was conducted to observe the electrophysiologic changes in C7 innervated muscles following transection of different fascicles of C7 nerve root in the brachial plexus with already transected C5 and C6. Twenty-four Sprague-Dawley rats were randomized into groups A, B, and C. C5 and C6 transection, C5, C6, and partial C7 (anteriolateral fascicles of the anterior division) transection, and C5, C6, and C7 transection were done in group A, B, and C, respectively. One month postoperatively, compound muscle action potentials of the latissimus dorsi, triceps, and extensor digitorum communis were recorded. Decreased amplitude and a prolonged latent period were seen in all three muscles of group C. No obvious changes, however, were found in groups A and B. This suggests that when the upper trunk of the brachial plexus is injured, partial transection of C7 does not obviously impair C7 innervated muscle function. Selective ipsilateral C7 transfer is safe for the treatment of upper-trunk avulsion.

[Construction of eukaryotic expression vector for rat myogenin gene].

OBJECTIVE: To construct eukaryotic expression vector of rat myogenin gene for further study on its functions in skeletal muscle denervated atrophy and repair. METHODS: The cloning vectors (containing full length of myogenin cDNA and two restriction sites: Hind III and Xho I) were first cut by two restriction endonuclease: Hind III and Xho I, and the same as the eukaryotic expression vector; then, the myogenin cDNA and the digested vector were ligated by T4 DNA ligase, and recombinant eukaryotic expression vector was formed. Its length was certificated by agarose gel electrophoresis analysis, digestion with Hind III and Xho I, PCR; and the rightness of the myogenin cDNA sequence was confirmed by sequencing. RESULTS: The results of agarose gel electrophoresis analysis, digestion, and PCR confirmed the right length of inserted DNA, which was the same as the myogenin cDNA, and the sequencing result of pcDNA3-myogenin was identical with the reported. CONCLUSION: pcDNA3-myogenin a eukaryotic expression vector, is successfully constructed.

Applying transfer of trapezius and/or latissimus dorsi with teres major for reconstruction of abduction and external rotation of the shoulder in obstetrical brachial plexus palsy.

To evaluate the effects of transfer of the trapezius and/or latissimus dorsi with the teres major for treatment of dysfunction of the shoulder in obstetrical brachial plexus palsy (OBPP), 34 patients with paresis of the abductors and external rotators, as well as co-contraction of the adductors in abduction, who had undergone reconstructive operations, were followed-up for at least 1 year. Of these, transfer of the latissimus dorsi with attached teres major to the insertion of the infraspinatus (single procedure), was performed in 25 cases, and transfer of both latissimus dorsi with teres major and trapezius (to the humerus) in nine (combined procedure). Gilbert's grading system was used for evaluation. The results showed that in spite of improvement of external rotation in most of the cases, abduction was improved in only 13 of the 25 cases with a single procedure, and that eight of nine cases with a combined procedure gained improvement of both external rotation and abduction. These results indicated that, for improvement of both abduction and external rotation of the shoulder in OBPP, transfer of the latissimus dorsi with the teres major can be performed only when abduction is > or =90 degrees; otherwise, transfer of the trapezius should be added.

[Ultrastructural changes after denervation of different muscles].

OBJECTIVE: To observe the ultrastructural changes and number of satellite cells in different muscles with different denervation interval and investigate the mechanism of denervation atrophy. METHODS: Muscles of different denervation interval were harvested, which were 6 biceps brachii and 6 abductor digiti minimi. The ultrastructure of the samples were observed under transmission electron microscope. The number of nucleus and satellite cells were counted to calculate the percentage content of satellite cells. RESULTS: In early stage of denervation, the myofilament and sarcomere of the majority were well oriented. The nucleoli of some muscle cell nucleus were enlarged and pale. Vacuolarization was also seen in some mitochondria. There was no obvious proliferation of collagen fiber around myofibers. After denervation of half a year, rupture and disorientation of myofilament was seen. The nucleus became smaller, dark stained, and some of them were condensed. There was proliferation of fibroblasts, adipose cells and collagen fibers around myofibers. Motor endplate was not recognized one year after denervation. In the early stage of denervation, satellite cell percentage of the two muscles was relatively high. It then declined with time. One year after denervation, satellite cells were scarcely detected. Comparison of the curves for satellite cell declination in two muscles revealed that the declination of the abductor digiti minimi was faster than that of biceps brachii. Decrease of the former started 3 months after denervation, while the latter started after 6 months. CONCLUSION: Disappearing of motor endplate and proliferation of collagen fibers are main factors that affect the treatment outcome in late cases. Decrease of satellite cell number is another cause. The correlation of less satellite cell in abductor digiti minimi and poorer recovery of hand intrinsic muscles indicates that increment of satellite cells in long-term denervated muscles may be one of the effective measures to improve treatment outcome.