Objective Bone marrow mesenchymal stem cells (BMSCs) were induced to differentiate to the special histological types of neurons in vitro.The morphological change of cells and positive expression of specific antigen on membrane were studied,and the function of connection between the induced BMSCs was also detected.The feasibility of BMSCs differentiate to the special histological types of neurons was investigated.Methods BMSCs were divided into group Ⅰ (induced with bFGF+GDNF),group Ⅱ (induced with bFGF+GDNF+WHI-P131 +Shh),and control group (no revulsive).The morphologic change of cells was observed,and the positive rate of neuron specific surface antigen and the content of dopamine were detected.Formation of mature synaptic structure was detected by immunohistochemical assay of postsynaptic density protein 95 (PSD-95) expression,and synaptic loop was shown by FM1-43 stain synaptic vesicles.Results By immunohistochemical staining,the positive rates of dopamine transporter (DAT) and tyrosine hydroxylase (TH) in group Ⅱ were significantly higher than those in group Ⅰ,and dopamine can been detected in cell culture supematant of group Ⅱ.After BMSCs was induced into dopamine neuron-like cells,number and length of cell protrusions,positive rate of PSD-95 and fluorescence intensity of FM1-43 in group Ⅱ were significantly higher than those of group Ⅰ.Conclusions There were no significant change in positive rate of neuron-specific surface markers,rate of cell survival and differentiation rate after BMSCs differentiated to dopaminergic neuron-like cells.The number and length of cell protrusions,content of dopamine in cell culture supematant,positive rate of dopaminergic neuron-specific surface antigen (DAT and TH),synaptic function index (positive rate of PSD-95 and fluorescence intensity of synaptic loop) of group Ⅱ were all significantly higher than that of group Ⅰ.

BACKGROUND: Pathophysiological mechanism of local microenvironment is complex after central nerve injury; especially,both inflammatory reaction at an acute phase and formation of secondary glial scar have tremendous effects on effective regeneration of axon, regeneration and arrangement of local nerve cells, proliferation and migration of local stem cells;therefore, it becomes a basic reason for blocking nerve repair in an early period. Thus, how to effectively resist inflammatory factors in injured region at an acute phase and how to optimize microenvironment of neural regeneration are the most important strategies for repairing spinal cord injury in recent years.OBJECTIVE: To design, establish and screen the best expression of interleukin-6 receptor (IL-6R) α to inhibit shRNA adenovirus expression vector by using spinal cord injury models.DESIGN: Duplicative measurement study.SETTING: Department of Spine Surgery, the Second People's Hospital of Shenzhen.MATERIALS: A total of 40 healthy Wistar rats, either gender, 8-10 weeks old, were selected in this study. Rabbit-anti-rat glial fibrillary acidic protein (GFAP) antibody Ⅰ was provided by Santa Cruz Compan; siRNA eukaryon expression plasmid pGenesil (cohtaining green fluorescent expression system) was provided by Wuhan Jingsai Bioengineering Company.METHODS: The experiment was carried out in ImmuneOpening Laboratory, Basic Medical Faulty, Tongji Medical College, Huazhong University of Science and Technology, and Medica Laboratory Center, the Second People's Hospital of Shenzhen in November 2006. Three pairs of shRNA template which composed of 19 bp reverse repeated motif of IL-6 receptor (IL-6R) α target sequence with 9 bp spacer were designed and synthesized, then the recombinant adenovirus expression vectors with green fluorescence protein were constructed in vitro respectively. The acute spinal cord injury models were completed, and the adenovirus recombinants were regionally injected post 12 hours after spinal cord injury;in addition, the inhibitory effect of RNA interference (RNAi) on expression of IL-6R in local region after spinal cord injury were detected by using real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot so as to screen adenovirus expression vector which had the best inhibitory effect on expression of IL-6R.MAIN OUTCOME MEASURES: Inhibitory effect of RNAi on expressions of IL-6R RNA and protein in local region after spinal cord injury.RESULTS: Sequence analysis showed that IL-6R-shRNA recombinant adenovirus expression vector was successfully constructed, and optimal IL-6R-shRNA recombinant adenovirus expression vector was screened by using real-time fluorescence quantitative RT-PCR and Western blot. The IL-6R expressions were 49% and 56% at the levels of mRNA and protein, respectively.CONCLUSION: The IL-6R--shRNA recombinant adenovirus expression vector is successfully constructed and screened.The gene expression of IL-6R can be highly inhibited after acute spinal cord injury.

Objective To produce a kind of biliary stent with Mitomycin C-eliting and evaluate the availability and safety in biliary benign stricture model of rabbit.Methods 36 New Zealand rabbits were fulgurize choledochus to establish model of biliary benign stricture.Rabbits were randomly divided into Mitomycin C-eluting stent group 1 (n =12),polyurethane stent group 2 (n =12) and control group 3 (n =12) one month later.General conditions,survival of the animals and changes in liver function were observed after surgery.The histological changes of bile duct were observed after 30 days.The immunohistochemistry SP method was used to measure transforming growth factor-β1 (TGF-β1) and α-Smooth muscle actin (α-SMA) expression.Results Stricture was improved in the two stent groups.In Mitomycin C-Eluting stent group total bilirubin dropped from 5.56 μmol/l to 0.82 μmol/l,and in polyurethane stent group total bilirubin dropped from 6.72 μmol/1 to 0.87 μmol/l.The total bilirubin decreased in both two stent groups but no statistically significant between the two stent groups,and there were no improvement in control group.Diameter of the stricture bile duct in group 1 was expanded bigger than in group 2 according to histology observation.Inflammatory cell infiltration and collagen fibroplasia in the submucosal were obviously observed in control group.The immunohistochemistry results showed that the TGF-β1 and α-SMA strongly expressed in the stenosis bile duct of group 3.The expressions in group 2 were lower than group 3,but higher than in group 1.And there was significant difference between the two stent groups (P ＜ 0.05).Conclusion The new Mitomycin C-Eluting stent is safe and provides enhanced local drug delivery.It also can inhibit the form of Biliary scar to a certain degree.

Fructus psoralea is a tonic traditional Chinese herb commonly used in clinic.The chemical constituents of Fructus psoralea are complicated,mainly containing coumarins,flavonoids and monoterpene phenols with various pharmacological effects.Since the increasing number of reports on the toxicity of Fructus psoralea in clinic,the side effects including toxicity on the liver and kidney,as well as skin allergies have gradually attracted attention.The toxicity of Fructus psoralea is produced from ADME (absorption,distribution,metabolism and excretion) in vivo.In this paper,we collected and clarified the studies of ADME of Fructus psoralea in vivo,and summarized recent adverse clinical events and research over its toxicity.We propose to make a thorough study on the toxic material basis of Fructus psoralea and the toxicological mechanism of its extraction,fractions and compounds.The review provided a possible reference and the direction of research for the safe clinical use of Fructus psoralea.

Objective To investigate the effect of medical rescue of the Maritime Medical Team (Corps) for mass sick and wounded in maritime disaster so as to improve the medical rescue capacity for maritime disasters.Method The construction of maritime medical teams (corps) constituted with various numbers of 10, 15,50 and 120 team members, and the development of algorithm in practice were reviewed. In 68 maritime disasters from January 2003 to December 2009, 937 wounded were rescued by first-aid at sea. The patients were classified and given cardiopulmonary cerebral resuscitation, emergency operation, complication prevention, comprehensive treatment for seawater immersion wound and rapidly referred to hospitals. Results Of 937 patients, 872 survived (93%) and 65 died (7%). Of the dead, 16 died in one hour (25%), 43 died in 24 hours after injury (66%),andofthem, 61died of trauma (94% ) , 2 died of drowning and 1 died of poisoning. Conclusions Besides a good command of the features of mass sick and wounded, organization and program, treatment strategies and measures, the timely and effective assignment for on-site first aid at sea and safe transfer were very important for medical rescue of mass patients in maritime disaster. After the practice of maritime medical team (corps) in medical rescue during maritime disaster, the rapid response capability, cooperation and the quality of rescue were improved, and the experience of medical service of marine medical team (corps) was enriched.

Clinical application of staphylococcal enterotoxin C2 (SEC2) was restricted during the cure of malignant tumor due to its side-effects. The aim of this study was to obtain SEC2 mutant, preserving the important functional sites responsible for the T-cell stimulatory activities but removing the sites responsible for emetic activity, through truncation of SEC2. It would efficiently solve the question of SEC2 side-effect. According to the results of methyl thiazol tetrazolium (MTT) assay in vitro, novel truncated SEC2 mutant (NSM) efficiently stimulated T-cell proliferation and inhibited the growth of such tumor cells as human colorectal cancer cells (Cx-1) and human breast cancer cells (MCF-7) in vitro. Activities of T cell stimulating and anti-tumor of NSM were similar to those of SEC2. According to results of animal experiments, the mutant no longer induced emetic response even if the dose was a 10-fold excess of the amount of SEC2 required. And also, NSM obviously inhibited the tumor growth in tumor-bearing mice. Therefore, we obtained novel truncated staphylococcal enterotoxin C2 mutant, which could efficiently inhibit the growth of tumor cells. It will become novel anti-tumor agents with the lowest side-effects and best treatment effects in clinic.
Animals ; Antineoplastic Agents ; adverse effects ; pharmacology ; Breast Neoplasms ; immunology ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; immunology ; pathology ; Enterotoxins ; genetics ; immunology ; Humans ; Mice ; Mutant Proteins ; immunology ; Staphylococcus aureus ; immunology ; Superantigens ; immunology ; T-Lymphocytes ; immunology ; Vomiting ; prevention & control

Objective To investigate the normal development of anterior and posterior acetabulum in children through measuring anterior and posterior acetabular indexes with the baseline from the thinnest point of acetabulum to the center of femoral head.Methods MRI data in 165 normal children aged 0-12 years were collected.The baselines were drawn from the center of the femoral head to the thinnest point of acetabulum (method 1) and from the one midpoint of Y cartilage to the contralateral (method 2),then the anterior or posterior bony acetabular index (A/PBAI) and anterior or posterior cartilaginous acetabular indexes (A/PCAI) were measured.The consistency of above parameters measured using two methods and between two observers was observed,and the correlation with parameters-gestational ages was analyzed.Results The consistency of ABAI (ICC=0.832) measured with two methods was good,and the consistency of ACAI (ICC=0.535),PBAI (ICC=0.565) and PCAI (ICC=0.472) was fair.The consistency between two observers was good (all ICC＞0.75).ABAI,ACAI and PBAI were negatively correlated with age (r=-0.762,-0.475,-0.368,all P＜0.001),and PCAI had no correlation with age (r=-0.190,P＜0.005).Before 4 years old,ABAI gradually decreased with age and gradually stabilized after 4 years of age.ACAI and PBAI decreased slightly with aging.PCAI did not change obviously with aging.Conclusion The measuring method of anterior and posterior acetabular indexes with the baseline from the thinnest point of acetabulum to the center of femoral head can accurately evaluate the normal development of anterior and posterior acetabulum in children.

Objective: To compare the effect of different orthokeratology lenses in controlling the progression of low myopia in children, and to provide a reference for exploring effective prevention measures for eyesight of children. Methods: A total of 175 cases (350 eyes) aged 8-12 years old who were fitted with orthokeratology lenses were collected in this retrospective study. The differences in the changes of the axis length (AL) and the spherical equivalent refraction (SER) were analyzed after wearing different orthokeratology lenses for one year, and the relationship between the change of AL, SER and gender, age was also analyzed. Results: In the Mouldway group, Alpha group, Lucid group and CRT group, the Median ( P 25 , P 75 ) of AL changes were 0.23 ( 0.12 , 0.41), 0.30 (0.17, 0.45), 0.35 (0.16, 0.41) and 0.33 (0.23, 0.41)mm, and there were no statistical significant difference between four groups ( Z =7.70, P >0.05); The Median ( P 25 , P 75 ) of SER changes were -0.31 (-1.00, 0.28), -0.38 ( -1.22 , 0.13), -0.25 (-0.84, 0.13) and -0.63 (-1.13, 0.25)D, and there were no statistical significant difference between four groups ( Z =2.15, P >0.05). The age had negative correlation with the change of AL ( r =-0.26, P <0.05), but has nothing to do with the change of SER ( r =0.10, P >0.05). There was no statistically significant difference in the change of AL ( Z =2.25, P > 0.05 ) and SER ( Z =-1.50, P >0.05) among children of different genders. Conclusion Different orthokeratology lenses have no differences in controlling the growth of the AL and changing the SER.

OBJECTIVETo investigate the value of C-reactive protein (CRP) on transplantation day in predicting early post-transplant infections and outcomes of allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSWe retrospectively analyzed the clinical data of 78 recipients undergoing allo-HSCT. The clinical reference value of CRP on transplantation day was determined, and its sensitivity and specificity for diagnosing bacteremia was analyzed using receiver-operating characteristic curve (ROC). The incidence of transplant-related complications, overall survival, and relapse rate of the patients were analyzed with respect to the CRP level.

RESULTSThe clinical reference value of CRP for diagnosing bacteremia was 23.3 mg/L (AUC=0.735 [95% CI: 0.623-0.848], P=0.001), which had a diagnostic sensitivity and specificity of 0.793 and 0.592, respectively. Compared with the patients with low CRP levels, the patients with high CRP levels tended to have delayed neutrophil reconstitution and platelet engraftment by 0.71 days (P=0.237) and 4.09 days (P=0.048), respectively, and had a significantly higher incidence of bacteremia (17.1% vs 53.5%, P=0.001) and CMV viremia (37.1% vs 72.1%, P=0.003) within 100 days following the transplantation; the incidences of EBV viremia, pulmonary invasive fungal infection, or acute graft versus host disease (aGVHD) showed no significant difference between the two groups (41.9% vs 22.9%, P=0.094; 14.0% vs 5.7%, P=0.285; 51.2% vs 45.7, P=0.656, respectively). During the follow-up for a median of 318 (7-773) days in high-CRP group and for 299 (78-747) days in low-CRP group, the high-CRP group showed a significantly lower 2-year overall survival than the low-CRP group (42.5% vs 78.4%, P=0.022), and tended to have a higher 2-year cumulative relapse rate (52.3% vs 19.8%, P=0.235). Logistic multivariate analysis identified a high CRP level on transplantation day as the independent risk factor for post-transplant bacteremia within 100 days (OR=5.090 [95% CI: 1.115 -23.229], P=0.036).

CONCLUSIONA high CRP level on transplantation day can be indicative of a high risk of early post-transplant bacteremia and CMV viremia and also a poor prognosis following allo-HSCT.

Bacteremia ; diagnosis ; C-Reactive Protein ; chemistry ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Incidence ; Mycoses ; Prognosis ; Recurrence ; Retrospective Studies ; Risk Factors ; Sensitivity and Specificity ; Viremia ; diagnosis

ObjectiveTo explore the mechanism of Chaihu Guizhitang （CHGZT） in alleviating neuropathic abdominal pain induced by 2，4，6-trinitrobenzene sulfonic acid （TNBS） in rats with chronic pancreatitis （CP）. MethodFifty male SD rats were randomly assigned into five groups： sham operation， CP model， and low-， medium-， and high-dose （4， 8， and 16 g·kg^-1， respectively） CHGZT groups. In the sham operation group， the abdomen was closed after the pancreas was gently stirred. The rat model of CP was established by retrograde injection of 2% TNBS-10% ethanol into the pancreatic duct. The oral administration of CHGZT started 4 weeks after modeling and lasted for 2 weeks. Pain threshold was measured by Von Frey fibers 6 weeks after surgery. Hematoxylin-eosin （HE） staining was employed to reveal the chronic inflammation and fibrosis of the pancreatic tissue. Immunohistochemmistry （IHC） was employed to detect the expression of PGP9.5 （a marker of pancreatic nerves） and reveal the inflammatory changes around the nerves. IHC and immunofluorescence （IF） were used to determine the location of ionized calcium-binding adaptor molecule-1 （Iba-1， microglia marker） and purinergic receptor P2X7 （P2RX7） and the co-expression of P2RX7 and Iba-1 in the thoracic spinal dorsal horn. ResultCompared with the sham operation group， the modeling increased the scores of pancreatic gland atrophy， inflammatory infiltration， and fibrosis （P<0.01）， the abdominal pain response under different force values （P<0.05， P<0.01）， and the score of peripancreatic inflammation. Moreover， the modeling up-regulated the expression of Iba-1 and P2RX7 in the thoracic spinal dorsal horn （P<0.01）. Compared with the model group， the high- and medium-dose CHGZT lowered the scores of pancreatic gland atrophy， inflammatory infiltration， and fibrosis， the abdominal pain response， and the score of peripancreatic inflammation （P<0.05， P<0.01）. The high-， medium-， and low-dose CHGZT all down-regulated the expression of Iba-1 and P2RX7 （P<0.01）. ConclusionCHGZT can significantly relieve abdominal pain in CP rat by suppressing the inflammation around nerves in the pancreas and the P2RX7 activation of microglia in the spinal dorsal horn.