Objective This study is undertaken to evaluate the changes of serum cytokine levels in different stages of collagen induced arthritis(CIA)rats,to search for the specific proteins related with rheuma- toid arthritis(RA)pathogenesis and inflammation,and to explore the mechanism of RA pathogenesis.Methods Rat cytokine antibody array coated with 19 specific cytokine antibodies was used to examine serum samples at peak and late stage of CIA rats,and were compared to normal cytokine levels.At the same time,ELISA assay for serum TNF-?production was used to verify the array results.Results Among the target cytokines,10 up- regulating cytokines were kept in high expression in different phases of disease,while 1 showed significant change only at the peak of disease.There was no downregnlating cytokines in the results.Serum TNF-?assay results were consistent to the array results.Conclusion Cytokines show different expression in CIA at differ- ent stages,and specific cytokines can be used as the candidates to further study of the RA pathogenesis.This study also provides molecular makers for early diagnosis.

Objective To compare the clinical efficacies of 3D and 2D laparoscopic partial nephrectomy for renal tumors and to investigate the pros and cons of 3D laparoscopic partial nephrectomy. Methods We retrospectively analyzed the clinical data of 134 patients with renal tumors; they underwent laparoscopic partial nephrectomy between Aug. 2013 and Mar. 2016 by the same surgical group in Changhai Hospital and Changzheng Hospital, Second Military Medical University. Fifty-three cases were included in the 3D laparoscopic partial nephrectomy group and 81 cases in the 2D laparoscopic partial nephrectomy group. The perioperative and follow-up data subjected to comparison and analysis included operative time, estimated blood loss, intraoperative transfusion, warm ischemic time, recovery time of bowel function, length of hospitalization, perioperative complications, and pre- and post-operative glomerular filtration rate (GFR). Results All operation procedures of this study were completed uneventfully without conversion to open surgery or radical nephrectomy, and surgical margins were negative in all cases. The operative time ([193.40±45.14] vs [217.00±59.19] min, P = 0.015), warm ischemic time ([23.70±6.96] vs [26.60±8.10] min, P = 0.032) and decrease of GFR of the operated kidney ([12.70 + 6.49] vs [15. 10 + 6. 45] mL • min-1 • 1. 73 m2, P = 0. 036) in the 3D group were significantly improved compared with those in the 2D group. However, the estimated blood loss, intraoperative transfusion, recovery time of bowtt function, length of hospitalization and perioperative complications were not significantly different between the two groups (P>0. 05). Att the patients with complications were discharged from the hospitat after conservative therapy. During a follow-up of 1-32 months, no patients had acute or chronic renat failure, local recurrence or distant metastasis, and there was no death case. Conclusion Compared with 2D laparoscopic partial nephrectomy, 3D laparoscopic technology can reduce operative time and warm ischemic time, which can help to better conserve renal function.

Objective: To investigate the changes of gene expression in CD34+ hematopoietic stem and progenitor cells (HSPCs) under different growth environments. Methods: Umbilical cord blood mononuclear cells (UCB MNCs) were cultured in static and stirred systems. After 7 days of culture, CD34+ cells were isolated and total RNA was extracted. Gene expression patterns of CD34+ cells from fresh, static and stirred cultures were compared using differential display (DD). Results: 30 gene fragments displayed differential expression levels based on the conditions of DD. One of differentially expressed genes was identified as RAN, which is a member of oncogene RAS family. This gene may be associated with proliferation of hematopoietic cells. Conclusion: Different growth environments induced differential gene expression patterns of CD34+ HSPCs. These differentially expressed genes would give new insights into optimizing in vitro environments for expanding hematopoietic cells.

The effect of the calcium ion (Ca2+) on the growth and differentiation of the mouse epidermal keratinocytes cultured in serum-free medium was investigated. It was found that the optimal level of calcium ion in the medium was about 0.2 mmol/L. Under such a culture condition the colony forming efficiency, attachment percentage, percentage of the cells with cornified envelops, and percentage of the senesced cells were measured to be about 10.8%, 30.8%, 5.1%, and 26.8%, respectively. However, the Ca2+ concentrations in the medium above 0.6 mmol/L resulted in significant differentiation and senescence of the keratinocytes, which was found to be harmful for keratinocyte growth and expansion in vitro.
Animals ; Calcium ; pharmacology ; Cell Adhesion ; drug effects ; Cell Differentiation ; Cell Division ; drug effects ; Cells, Cultured ; Cellular Senescence ; drug effects ; Epidermis ; cytology ; Keratinocytes ; cytology ; drug effects ; Mice

OBJECTIVETo investigate the late results of using posterior restoration and three-column fixation to treat lumbar spondylolisthesis.

METHODSOne hundred and eighty-four patients with lumbar spondylolisthesis were collected from March 1999 to May 2007, they were treated by posterior restoration and fixation with single nail-grooved tail steel plate and fixed with cage (WDFC). Among these cases, 87 cases were fixed with one WDFC, 97 cases were used two WDFCs.

RESULTSAll patients were followed up for 8 to 69 months(averaged 23 months). According to Nakai standard, the results was excellent in 142 cases, good in 34, fair in 8, the excellent and good rates were 95.6%. Seventy-nine vertebraes with I degree spondylolisthesis were reduced after surgery. Eighty-seven vertebraes with II degree spondylolisthesis were reduced except 9 with I degree spondylolisthesis left. Twenty-one with III degree spondylolisthesis were reduced except 5 with I degree spondylolisthesis left; In 2 with IV degree spondylolisthesis, one with I degree spondylolisthesis left and the other with II degree spondylolisthesis left. The follow-up results showed that there was no statistical significance in the height of intervertebral space between preoperation and post-operation, and no recurrence was observed and no single nail-grooved tail steel plate and WDFC were loose or crashed.

CONCLUSIONPosterior restoration and three-column fixation is a positive modus operandi to treat lumbar spondylolisthesis,which can reduce excellently,keep the height of intervertebral space and stabilization of segment, obtain high rate of fusion, and cut down complication.

Adolescent ; Adult ; Aged ; Bone Plates ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; instrumentation ; Spondylolisthesis ; surgery

To optimize the culture environment and protocol of hematopoietic cells' expansion, avoiding the fluctuation caused by medium changing in stirred culture and concentration gradient in static culture, the hematopoietic cells from cord blood (CB) were cultured in a stirred bioreactor connected with a cell retention system, which is a gravity sedimentation settler designed for hematopoietic cell. Total cells expanded 11.5 and 18.6 fold respectively in the twice perfusion stirred cultures, in which CFU-Mix was expanded 23.2 and 20.4 fold, CFU-GM 13.9 fold and 21.5 fold, BFU-E 8.0 fold and 6.9 fold, CD34+ cells 17.1 fold and 15.4 fold. After 12-day culture, it was obtained that 1082 x 10(6) total cells, 6.31 x 10(6) CFU-GM, 6.2 x 10(6) CFU-Mix and 23 x 10(6) CD34+ cells from 267 x 10(6) CB mononuclear cells (MNC) in the first culture, and 1080 x 10(6) total cells, 4.65 x 10(6) CFU-GM, 11.0 x 10(6) CFU-Mix, and 25.0 x 10(6) CD34+ cells from 180 x 10(6) CB MNC. These two cultures met to the clinical scale. Due to the optimized dissolved oxygen (DO) and stable culture environment, the rate of stem/progenitor cells to total cells in the perfusion culture was higher than that in T-flask cell-retention feeding culture. But the cell growth was inhibited in the later phase of perfusion culture, when the cell density is high. The inhibition should be attribute to the high cell density itself. The perfusion culture environment in bioreactor with optimal DO and pH controlling is more favorable for stem/progenitor cells' maintenance and expansion, and the expanded cells' number has reached a clinical scale. But the high cell density in the later phase of perfusion culture caused inhibition to mature hematopoietic cell's growth.
Bioreactors ; Cell Culture Techniques ; methods ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans

OBJECTIVETo construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om.

METHODShCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting.

RESULTShCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 kD hCD96om fusion protein.

CONCLUSIONThe CD96 gene of human has been successfully cloned and expressed in BL21(DE3) cells, and its rabbit polyclonal antibody has been obtained.

Animals ; Antibodies ; immunology ; metabolism ; Antigens, CD ; biosynthesis ; genetics ; immunology ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Humans ; Immune Sera ; biosynthesis ; Immunization ; Leukemia, Myeloid, Acute ; immunology ; Molecular Sequence Data ; Neoplastic Stem Cells ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo study apoptosis and gene FasL expression induced by carbon disulfide in sertoli cells of male rats.

METHODSSertoli cells were exposed to different concentrations of CS(2) (0, 0.36, 0.72, 1.44 micromol/ml) for 24 hours. Survival rate, apoptosis rate, expression level of gene FasL were measured using MTT, FCM, and RT-PCR methods respectively.

RESULTSSertoli cell survival rate decreased as the concentration of CS(2) increased. The survival rate (73.34% +/- 1.39%) was significantly lower than the control group (99.98% +/- 5.48%) when the concentration of CS(2) > or = 1.44 micromol/ml (P < 0.05). Apoptosis rate increased as the CS(2) concentration increased. Apoptosis rate (7.93% +/- 0.43%) was significantly higher when the concentration of CS(2) > or = 1.44 micromol/ml (P < 0.05). Expression level of the FasL significantly increased as the concentrations of CS(2) (P < 0.05).

CONCLUSIONCS(2) is cytotoxic to sertoli cells. It could cause apoptosis of sertoli cells.

Animals ; Apoptosis ; drug effects ; Carbon Disulfide ; toxicity ; Cell Line ; Cell Survival ; Fas Ligand Protein ; metabolism ; Male ; Rats ; Sertoli Cells ; drug effects ; metabolism ; Testis ; cytology

Objective To construct a digital atlas of the globus pallidus internus (Gpi) in the standard stereotactic space using the sectional images of the male cadaver from "Atlas of Chinese Visible Human (Male and Female)". Methods Identification and segmentation of the Gpi,and anterior and posterior commissures from the sectional images were performed. We established the stereotactic coordinate system and reconstructed 3D models of the Gpi. Results Gpi looks like a bean according to the reconstructed models; Putamen and globus pallidus lateral (Gpe) were also constructed; and the location of the Gpi and the relation of Gpi with both Putamen and Gpe were shown in the stereotactic space. The lenticular nucleus could rotate and stretch at any angles in the standard stereotactic space.Conclusion It is feasible to reconstruct the 3D models of the subnucleus in the stereotactic space from the sectional image of the cadaver.

BACKGROUNDSpinocerebellar ataxia type 3 (SCA3) is the most common subtype of SCA worldwide, and runs a slowly progressive and unremitting disease course. There is currently no curable treatment available. Growing evidence has suggested that nerve growth factor (NGF) may have therapeutic effects in neurodegenerative diseases, and possibly also in SCA3. The objective of this study was to test the efficacy of NGF in SCA3 patients.

METHODSWe performed an open-label prospective study in genetically confirmed adult (>18 years old) SCA3 patients. NGF was administered by intramuscular injection (18 μg once daily) for 28 days consecutively. All the patients were evaluated at baseline and 2 and 4 weeks after treatment using the Chinese version of the scale for assessment and rating of ataxia (SARA).

RESULTSTwenty-one SCA3 patients (10 men and 11 women, mean age 39.14 ± 7.81 years, mean disease duration 4.14 ± 1.90 years, mean CAG repeats number 77.57 ± 2.27) were enrolled. After 28 days of NGF treatment, the mean total SARA score decreased significantly from a baseline of 8.48 ± 2.40 to 6.30 ± 1.87 (P < 0.001). Subsections SARA scores also showed significant improvements in stance (P = 0.003), speech (P = 0.023), finger chase (P = 0.015), fast alternating hand movements (P = 0.009), and heel-shin slide (P = 0.001).

CONCLUSIONSOur preliminary data suggest that NGF may be effective in treating patients with SCA3.

Adult ; Animals ; Female ; Humans ; Injections, Intramuscular ; Machado-Joseph Disease ; drug therapy ; Male ; Mice ; Middle Aged ; Nerve Growth Factor ; administration & dosage ; therapeutic use ; Prospective Studies