The aim of this study was to investigate if transfusion of mesenchymal stem cells (MSC) could exhibit beneficial effects on rheumatoid arthritis. Human bone marrow MSC were intraperitoneally injected into Wistar rats with collagen-induced arthritis at a dose of 10(7) on the next day (preventive group) or 2 weeks (treatment group) after collagen II induction, once a week for 2 weeks (preventive group) or 4 weeks (treatment group). The control group was given normal saline (NS) at corresponding time. The symptom scorings were documented weekly from the second week of the induction. On week 6, the hind joints of the rats were pathologically examined and the activation status of splenocytes was analyzed by flow cytometry. The results showed that all the rats developed arthritis and subsequent joint abnormality. On the sixth week, symptom scores of the rats that received MSC preventive (9.5 ± 0.5) or therapeutic (9.4 ± 0.6) infusions had no significant difference between each other, but were significantly greater than those of the NS controls (7.6 ± 0.6, P < 0.05). Consistently, pathological examination on the involved knees showed that the synovitis and arthritis scorings of MSC treated rats were greatly elevated compared with NS controls. Furthermore, the ratios of CD86(+) cells in the spleens of MSC prevention, MSC treatment and NS control groups were (4.16 ± 1.48), (4.06 ± 1.97) and (4.15 ± 2.04) respectively, while those of CD11b/c(+)CD86(+) cells were (1.04 ± 0.68), (0.95 ± 0.56) and (0.98 ± 0.44), all of which were significantly higher than those of healthy controls [(0.97 ± 0.18) and (0.30 ± 0.17), P < 0.05 for both parameters]. It is concluded that MSC infusion has little beneficial effects on collagen-induced arthritis in rats, conversely, MSC therapy aggravated the damage of the involved joints, its underlying mechanisms need to be further investigated.
Animals ; Arthritis, Experimental ; pathology ; prevention & control ; therapy ; Bone Marrow Cells ; Cells, Cultured ; Humans ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Wistar ; Transplantation, Heterologous ; Treatment Outcome

Mesenchymal stem cells (MSC) are characterized by their potent immuno-regulatory activity, however our previous data have shown that MSC have no therapeutic effects on collagen-induced arthritis (CIA). To further clarify the complexity, the effects of tumor necrosis factor-alpha (TNF-α) on the in vitro and in vivo immunoregulatory activity of MSC were investigated in this study, as TNF-α is recognized as the key factor in the development of rheumatoid arthritis. The nuclear translocation of the inflammation-associated factor NF-κB was observed after human umbilical cord MSC were treated with TNF-α and the cell proliferation status was assessed by MTT test. The inhibitory effects of MSC or TNF-α-treated MSC on the mixed lymphocyte reaction, in which Wistar rat spleen mononuclear cells were served as the responders and the splenocytes from SD rat spleens as the stimulators, were also determined by the MTT test. Further, the therapeutic potentials of MSC or TNF-α-treated MSC were observed in a Wistar rat CIA model. The results showed that NF-κB translocated into the nuclei promptly after TNF-α treatment, though TNF-α had little effect on the MSC proliferation. MSC, whether pre-stimulated by TNF-α or not and when different doses were tested, exhibited obviously inhibitory effects on the proliferation of the lymphocytes (P < 0.001 for all groups tested), while MSC-treated by TNF-α displayed more potent suppression especially when low-density were used. Unexpectedly, the infiltration of inflammatory cells into the involved knees was aggravated by cell treatment and the pathological scores were significantly higher than those of controls (P < 0.05). It is concluded that the TNF-α exhibits different effects on immune regulation activity of MSC, and its underlying mechanism needs to further investigate.
Animals ; Arthritis, Experimental ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; immunology ; NF-kappa B ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; pharmacology

This study was purposed to investigate the immune reconstitution of T-cells in patients who received haploidentical hematopoietic stem cell transplantation (hiHSCT). The peripheral blood was harvested from 22 patients before transplantation and at month 1, 3, 6 after hiHSCT. The proportions of T lymphocyte subtypes including CD3(+), CD4(+), CD8(+), CD45RO(+), and CD45RA(+)CD62L(+) were analyzed by flow cytometry, followed by the calculation of T cell numbers according to the amounts of peripheral blood leukocytes. Adenosine triphosphate (ATP) value in CD4(+) T cells was measured by ImmuKnow method to evaluate the function of lymphocytes. The results showed that the CD3(+) cell absolute value before transplantation was 833.75 ± 359.84/µl, but those values at month 1, 3, 6 after transplantation were 318.87 ± 266.71/µl, 1006.76 ± 512.32/µl and 1296.38 ± 958.77/µl respectively. The CD4(+) cell absolute value before transplantation was 336.99 ± 211.11/µl, but such values at month 1, 3, 6 after transplantation were 45.89 ± 44.21/µl, 142.97 ± 114.85/µl, and 181.78 ± 120.61/µl respectively. The CD8(+) cell absolute value before transplantation was 430.21 ± 159.48/µl, but those values at month 1, 3, 6 after transplantation were 230.44 ± 195.89/µl, 621.64 ± 318.83/µl, and 823.07 ± 633.55/µl respectively. The CD4(+)CD45RO(+) memory T cell absolute value before transplantation was 227.44 ± 73.34/µl, but such values at month 1, 3, 6 after transplantation were 43.47 ± 43.40/µl, 138.69 ± 110.17/µl, 147.73 ± 82.94/µl respectively. The CD8(+)CD45RO(+) memory T cell absolute value before transplantation was 212.70 ± 98.48/µl, but such values at month 1, 3, 6 after transplantation were 184.76 ± 168.65/µl, 445.90 ± 252.50/µl, 519.80 ± 475.53/µl respectively. CD4(+)CD45RA(+)CD62L(+) naive T cell number before transplantation was 68.94 ± 59.74/µl, but such cell numbers at month 1, 3, 6 after transplantation decreased to 2.44 ± 2.93/µl, 3.14 ± 3.48/µl, 23.22 ± 38.38/µl respectively. The CD8(+)CD45RA(+)CD62L(+) naive T cell absolute value before transplantation was 124.82 ± 60.95/µl, but those values at month 1, 3, 6 decreased to 19.37 ± 17.71/µl, 76.63 ± 50.85/µl, and 114.49 ± 174.29/µl respectively. The ATP value in CD4(+) T cells decreased to 210.19 ± 119.37 ng/ml at month 1 after transplantation and increased to 280.62 ± 110.03 ng/ml at month 3, and 357.28 ± 76.18 ng/ml at month 6 after transplantation. It is concluded that CD8(+) memory T cell reconstruction contributes critically to T cell recovery early after hiHSCT, while the thymic output function remains low. However, T cell function recovers to normal range at month 3 after transplantation.
Adolescent ; Adult ; CD8-Positive T-Lymphocytes ; cytology ; Child ; Child, Preschool ; Female ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Immunophenotyping ; Killer Cells, Natural ; immunology ; Lymphocyte Count ; Male ; T-Lymphocyte Subsets ; immunology ; Young Adult

Our previous work has shown that mesenchymal stem cells (MSC) have little therapeutic effect on rat arthritis induced by collagen. This study was aimed to further investigate whether the MSC lysates exhibit beneficial effects on rheumatoid arthritis. Aliquots of cell lysates from 1×10(7) human bone marrow MSC were intraperitoneally injected into collagen-induced arthritis (CIA) Wistar rats weekly for 4 consecutive weeks. Methotrexate at a dose of 1 mg/kg or normal saline was served as positive and negative controls respectively. On week 4 the symptom scores were recorded and the hind joints of the rats were pathologically examined and X-ray examination was performed. The results showed that on week 4, the symptom scores of the rats that received MSC lysates (6.87 ± 0.83) and MTX (6.44 ± 1.13) were significantly lower than that of control rats (7.33 ± 0.77, P < 0.01). Meanwhile, pathological examination on the involved ankle showed that the synovitis and arthritis scores of MSC lysates and control groups were 2.28 ± 0.48 and 2.28 ± 0.55 respectively, significantly higher than that of MTX treatment rats (0.71 ± 0.48, P < 0.05). However, X-ray examination on the ankle joints showed that the injury score of control rats was 4 ± 0.57, greatly higher than those from MSC lysates (2.71 ± 0.75) and MTX treatment groups (2.57 ± 0.78, P < 0.05 for both groups). It is concluded that MSC lysate infusion has beneficial effects on CIA rat, but the effectiveness seems inferior to MTX.
Animals ; Arthritis, Experimental ; chemically induced ; therapy ; Bone Marrow Cells ; cytology ; Cells, Cultured ; Collagen ; Humans ; Male ; Mesenchymal Stromal Cells ; cytology ; Methotrexate ; pharmacology ; Rats ; Rats, Wistar

OBJECTIVETo study processing method and mechanism of Calamine.

METHODThermogravimetry analysis method and nano-technology were adopted to analyze and synthesize the components in Calamine, Tetracycline was took as the comparison drug to determine the antibacterial activity of Calamine and its components.

RESULTA part of zinc carbonate in Calamine was decomposed into zinc oxide when processing, and the particle size was smaller than before. The antibacterial activity of Calamine is decided by the content and particle size of zinc oxide, and has nothing with zinc carbonate. The more content and the smaller particle size of zinc oxide, the more powerful antibacterial activity of Calamine.

CONCLUSIONThe content and the particle size of zinc oxide can be the important targets in the processing of Calamine.

Anti-Bacterial Agents ; pharmacology ; Carbonates ; chemistry ; pharmacology ; Drug Combinations ; Escherichia coli ; drug effects ; Ferric Compounds ; chemistry ; pharmacology ; Materia Medica ; chemistry ; pharmacology ; Nanostructures ; Nanotechnology ; Particle Size ; Pseudomonas aeruginosa ; drug effects ; Salmonella ; drug effects ; Staphylococcus aureus ; drug effects ; Technology, Pharmaceutical ; methods ; Tetracycline ; pharmacology ; Thermogravimetry ; Zinc Compounds ; chemistry ; pharmacology ; Zinc Oxide ; analysis ; chemistry ; pharmacology

Objective:To study the serological characteristics of ABO*A2.08 subtype and explore its genetic molecular mechanism.Methods:ABO blood group identification was performed on proband and her family members by routine serological methods.ABO genotyping and sequence analysis were performed by polymerase chain reaction-sequence specific primer(PCR-SSP),and direct sequencing of PCR products from exons 6 and 7 of ABO gene were directly sequenced and analyzed.The effect of gene mutation in A2.08 subtype on structural stability of GTA protein was investigated by homologous protein conserved analysis,3D molecular modeling and protein stability prediction.Results:The proband's serological test results showed subtype Ax,and ABO genotyping confirmed that the proband's genotype was ABO*A207/08.Gene sequencing of the proband's father confirmed the characteristic variation of c.539G＞C in the 7th exon of ABO gene,leading to the replacement of polypeptide chain p.Arg180Pro(R180P).3D protein molecular modeling and analysis suggested that the number of hydrogen bonds of local amino acids in the protein structure was changed after the mutation,and protein stability prediction showed that the mutation had a great influence on the protein structure stability.Conclusion:The mutation of the 7th exon c.539G＞C of ABO gene leads to the substitution of polypeptide chain amino acid,which affects the structural stability of GTA protein and leads to the change of enzyme activity,resulting in the A2.08 phenotype.The mutated gene can be stably inherited.

OBJECTIVETo evaluate the mid-term clinical outcomes of minimally invasive transforaminal lumbar interbody fusion (TLIF) with unilateral pedicle screw fixation for lower lumbar degenerative diseases.

METHODSFrom April 2004 to December 2005, minimally invasive TLIF through paramedian approach with unilateral pedicle screw fixation was performed in a consecutive series of 43 patients, including 24 male and 19 female, aging from 38 to 71 years, with an average age of 49 years. The length of surgical incision was 3 cm. The operation level at L(3-4) were 3 cases, L(4-5) 27 cases, L(5)-S(1) 13 cases and no case was at multilevel. Clinical outcomes were assessed by ODI scores and JOA questionnaires before and after operation. Operation time, intraoperative blood loss, incision status and complications were recorded. Radiological examination was obtained for each patient to assess the height of intervertebral space, postoperative intervertebral fusion conditions and the degeneration of adjacent segments.

RESULTSThe mean operation time was 110 minutes, the mean blood loss was 150 ml and all the incisions were healed primarily. The follow-up time ranged from 36 to 58 months. The ODI scores decreased significantly from 60 ± 10 preoperatively to 12 ± 4 postoperatively (P < 0.01). The JOA scores were improved remarkably from 9.6 ± 2.2 preoperatively to 23.8 ± 2.0 postoperatively (P < 0.01) and the proportion with optimal effect was 86%. The ventral and dorsal heights of intervertebral disc were significantly higher than those before operation (P < 0.01). The fusion rate was 94%. The incidence of adjacent segment degeneration was 17%. There were no complications such as secondary scoliosis, screw loosening, internal fixation failure and cage slippage.

CONCLUSIONSThe minimally invasive TLIF through paramedian approach with unilateral pedicle screw fixation is an effective and convenient method with little surgical trauma. The mid-term follow up results showed favorable outcomes in patients receiving this surgery.

Adult ; Aged ; Bone Screws ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; methods ; Spondylolisthesis ; etiology ; surgery ; Treatment Outcome

Etanercept has been shown to be effective for the treatment of moderate-to-severe plaque psoriasis.Since most clinical trials examined etanercept in combination with other drugs,the efficacy and safety of etanercept monotherapy for moderate-to-severe plaque psoriasis have not been well established.This prospective study enrolled 61 Chinese patients with moderate-to-severe plaque psoriasis to explore the efficacy and safety of etanercept monotherapy.These patients were treated with etanercept at a subcutaneous dose of 25 mg,twice a week,for 12 weeks.All the 61 patients completed the treatment and showed significant improvement in psoriasis area and severity index (PASI) scores.At 4,8,and 12 weeks after treatment,the response rates (PASI75) were 0％,21.31％,and 40.98％,respectively.It was concluded that etanercept monotherapy is efficacious and safe for patients with moderate-to-severe plaque psoriasis.

OBJECTIVETo assess the clinical outcomes of Coflex interspinous dynamic internal fixation and the imaging changes for degenerative lumbar spinal stenosis.

METHODSFrom October 2007 to February 2009, 30 patients with degenerative lumbar spinal stenosis were treated with Coflex interspinous dynamic internal fixation, including 17 males and 13 females with an average age of 45 years (range, 39 to 65 years). The operation level at L4,5 were 20 cases, L5S1 9 cases and 1 case was in both the two levels. The ODI scores and JOA questionnaires were assessed before and after operation. The radiological measurement included ventral and dorsal intervertebral space height, segmental intervertebral angles formed by lines drawn on the upper and lower endplates of the instrumented and adjacent levels on flexion-extension radiographs; the area of spinal canal, dural sac, the sagittal and transverse diameter of the spinal canal and dural sac on CT scan. All the patients were treated with limited laminectomy and were implanted with Coflex device.

RESULTSAll the patients were followed up for 5 to 19 months. There were significant differences in the ODI scores and the JOA questionnaires by paired t-test (P<0.01). All the patients were satisfied with surgical outcomes except 3 patients whose pain were not obviously relieved and need drugs or block therapy; 3 patient complaining of progressive hypoesthesia. There were no complications associated with the Coflex device. The height of dorso-intervertebral disc was increased obviously while the range of motion in adjacent levels was not increased on flexion-extension radiographs; the area of spinal canal and dural sac were significantly increased.

CONCLUSIONIt shows a good clinical result to release the degenerative lumbar spinal stenosis symptoms and decrease short-term complications by using Coflex device. It is available for patients with increasing the intervertebral space, area of spinal canal and preventing the adjacent segment degeneration.

Adult ; Aged ; Decompression, Surgical ; methods ; Female ; Humans ; Internal Fixators ; Lumbar Vertebrae ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Radiography ; Spinal Stenosis ; diagnostic imaging ; surgery ; Treatment Outcome

OBJECTIVETo compare clinical efficacy between discectomy and discectomy plus Coflex fixation for lumbar disc herniation.

METHODSFrom December 2007 to August 2008, 50 patients (31 males and 19 females) were treated by surgery of discectomy and discectomy plus Coflex fixation. The average age was 52.5 years (range, 30 - 72 years). There were 24 cases in the group of discectomy plus Coflex fixation and 26 cases in the group of discectomy. Preoperative and postoperative visual analogue scales (VAS), Japanese Orthopadic Association (JOA) and Oswestry disability index (ODI) were recorded, as well as radiological index. And use a paired t-test and one-way analysis of variance (one-way ANOVA) statistical method to evaluate the Coflex dynamic stabilization system in value in the treatment of lumbar disc herniation.

RESULTSBoth groups received significant improvement of JOA, ODI and VAS (t = -33.2 - 64.5, P < 0.01), but the group of discectomy was found with deterioration of ODI at last follow-up, 12 months after surgery 6.7 ± 1.5 to 10.2 ± 2.3 (t = -19.3, P < 0.05). The group of discectomy plus Coflex fixation was found with significant increase of height of dorsal intervertebral discs (HD), distance across the two adjacent spinous processes (DS), distance of intervertebral foramina (DIF) and spinal canal area(SA) (t = -34.4 - 4.5, P < 0.05). In contrast, the group of discectomy was found with significant decrease of HD, DS, DIF and SA (t = 3.4 - 52.8, P < 0.05). Coflex fixed group in HD, DIF, DS significant difference with simple discectomy group, with a statistically significant (F = 14.1 - 25.6, P < 0.05).

CONCLUSIONSBoth discectomy and discectomy plus Coflex fixation are apparently effective when treating lumbar disc herniation. Coflex can significantly increase the HD and DIF when used for lumbar disc herniation, and it has positive influence for keeping height of lumbar vertebral space and treating the nerve root symptom of lumbar disc herniation. Discectomy plus Coflex is better than pure discectomy in preventing lumbar degeneration.

Adult ; Aged ; Female ; Humans ; Internal Fixators ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Treatment Outcome