BACKGROUND:Studies have shown that bone marrow mesenchymal stem cells under certain conditions can differentiate into nerve cells. Bone marrow mesenchymal stem celltransplantation can rebuild nervous system function and improve functional disorders in patients. Glycosides of cistanche also have a protective effect against nerve cellinjury. Their combination has been less reported. OBJECTIVE:To observe the effectiveness of Cistanche deserticola and bone marrow mesenchymal stem celltransplantation on spinal cord injury in rats. METHODS:Fifty adult Wistar rats with spinal cord injury were randomly divided into four group:a Cistanche deserticola group (intragastric administration of 20 mL/kg Cistanche deserticola concentrated solution per day), a celltransplantation group (10μL of 1×108/L bone marrow mesenchymal stem cellsuspension), a combination group (10μL of 1×108/L bone marrow mesenchymal stem cellsuspension+intragastric administration of 20 mL/kg Cistanche deserticola concentrated solution per day) and a control group (intragastric administration of 20 mL/kg normal saline per day). The intragastric administration lasted for 30 days in each group. RESULTS AND CONCLUSION:After 30 days of treatment, the expression of Nestin was significantly higher in the combination group than the other groups. After 12 weeks, Basso, Beattie, and Bresnahan scores was significantly higher in the combination group than the other groups (P<0.05);somatosensory and motor evoked potential latencies were also improve significantly in the combination group compared with the other groups (P<0.05). These findings indicate that oral administration of Cistanche deserticola combined with bone marrow mesenchymal stem cells can significantly improve the motor and neurophysiological function of spinal cord injury rats. Cistanche deserticola can improve the survival of transplanted bone marrow mesenchymal stem cells in rats with spinal cord injury.

Objective To investigate the effect of EST on function of sphincter of Oddi and on the long-term complications.Methods A total of 68 patients with common bile duct stones were recruited from January 2009 to January 2013 in The First People's Hospital of Hangzhou.All patients were under follow-up to observe the incidence of late complications after EST.Oddi sphincter manometry and bile bacterial culture were performed before EST and one year later.Results With the 13-36 months follow-up,the incidence of late complications was 16.9％ (11/65).The peak pressure and frequency of contraction of Oddi sphincter after EST were significantly lower than those before EST,but there were no significant differences between patients with complication and without (P ＞ 0.05).After EST,positive rate of biliary bacteria in patients with complications was much higher than that in patients without complications [70％ (7/10)vs.31.7％ (13/41),P =0.026],although there were no significant differences between them before EST [54.5％(6/11) vs.46.3％ (25/54),P =0.618).Logistic regression analysis showed that cholecystolithiasis,bile duct diameter (＞ 1.5 cm),number of stone (＞ 3),maximun diameter of stone (＞ 20 mm),large EST were independent risk factors for late complications of EST.Conclusion Importance should be attached to the late complications of EST with Oddi sphincter function declination,and biliary enteric reflux being the basic factor in late compilations after EST,which are also influenced by many risk factors.

BACKGROUND:With the development of spinal cord injury study, different methods of establishing spinal cord injury models have emerged, including spinal cord contusion, fal ing weight, spinal compression, chemical burn, radiation, hormone, spinal transection and hemi-section. However, lots of them are not perfect enough.
OBJECTIVE:To design the injury device of spinal cord injury and establish different degrees of spinal cord injury models.
METHODS:To design the device of producing spinal cord injury and establish different degrees of spinal cord dorsal compression injury in Sprague-Dawley rats by various weights (m1=10 g, m2=20 g, m3=30 g) and time points (T1=3 s, T2=5 s). Rats were randomly divided as m1T1, m2T1, m3T1, m1T2, m2T2 and m3T2 groups. While sham group was also made.
RESULTS AND CONCLUSION:Basso-Beattie-Bresnahan (BBB) score in injury groups decreased significantly after operation, when compared with the sham group (P<0.01). The m 1T1 group showed no significant difference in BBB score from other groups (P<0.01). The BBB score of m1T2 group was significant higher than m2T2 group and m3T2 group at 8 weeks after operation (P<0.05). The somatosensory evoked potential and motion evoked potential of injury groups were longer than sham group at 8 weeks after operation (P<0.01). The motion evoked potential of each injury groups were significantly longer after operation (P<0.05). The somatosensory evoked potential was significantly longer in injury groups, except m1T1 and m1T2 groups (P<0.05). The self-designed device can be applied to establish different degrees of spinal cord injury models.

BACKGROUND:Total knee replacement is regarded an effective treatment of knee osteoarthritis, based on the transformation of biology-psychology-social medicine pattern, the concept of“quality of life”has attracted increasing concerns.
OBJECTIVE:To observe the effects of total knee replacement surgery on the pain, function and quality of life among elder patients with severe knee osteoarthritis.
METHODA total of 102 elder patients with severe knee osteoarthritis were recruited from the Second Affiliated Hospital of Inner Mongolia Medical University between January 2010 and January 2013. The clinical manifestations and 12-month fol ow-up outcomes were observed and recorded. The involved patients were randomly divided into two grouptreatment group (59 cases, receiving total knee replacement) and control group (43 cases, receiving expectant treatment such as drugs, but no surgery). Patients were evaluated using HSS scale and WHOQOL-100 scale before surgery and 12 months after surgery. Furthermore, the correlation between influencing factors and HSS and WHOQOL-100 scores was analyzed.
RESULTS AND CONCLUSION:At 12 months postoperatively, the HSS scores of patients in treatment group was (82.03±10.17) points, which was obviously higher than that in control group (P<0.01). The WHOQOL-100 scores in treatment group were also significantly higher than that in control group (P<0.01). The psychology sub-item scores of patients after total knee replacement were close to that of normal controls (P>0.50). The regression analysis shows that marital status (β=1.988), complication (β=2.035) and HSS scores (β=2.108) are the main influencing factors for the quality of life among patients undergoing replacement. Experimental findings indicate that, HSS score is an effective indicator to assess the effect of total knee replacement on the pain and function of elder patients with severe knee osteoarthritis. WHOQOL-100 is introduced to further improve the assessment of quality of life after total knee replacement surgery in severe knee osteoarthritis. The marital status, complication and HSS scores are the key influencing factors for the quality of life among severe knee osteoarthritis patients. Deep vein thrombosis affects the out-of-bed time, and the mate-accompanying during the fol ow-ups plays a positive role on the quality of life and mental state of patients.

Objective To explore for the methede and effect of endoscopic treatment on biliary leakage and biliary duct damage. Methods All patients with biliary damage such as biliary leakage and biliary duct stricture were treated by endoscopic sphincoterotomy and endoscopic nasobiliary drainage (ENBD) during abdominal cavity drainage ENBD was removd when biliary leakage healed and abdominal cavity drainage ceased for 1～2 weeks were confirmed. Plastic stents were implanted to distend the biliary duct stricture for 2-3 months. Results Twenty-six patients with biliary leakage were cured 3-4 weeks after ENBD. Fourteen out of 17 patients implanted with plastic stent were recovered uneventfully after stent removed, and 4 patients also recovered after installation of double-stents for 3 months, while another case with calculus and stricture of left hepatic duct in spite of implantation of simple-stent suffered repeatedly from biliary tract infection and one case developed hepatic abscess after repeatedly infection for one year before he had the hepatic lobectomy. Conclution Endoscopic therapy is the first choice in treating biliary leakage or secondary duct stricture.

Objective To evaluate brush cytology under endoscopic retrograde cholangiopancreatography (ERCP) for malignant biliary strictures, and to analyze the factors influencing the diagnosis yield.Methods Brush cytology was applied in 144 patients with suspected malignant biliary strictures at ERCP.Brushing in bile duct was performed for 10 times from 2004 to 2006, while at least 20 times of brushing and repeated manipulation were performed at biliary strictures from 2007 to 2009. Cytological samples were processed immediately after brushing and analyzed by the same pathologist. Efficacy of brush diagnosis was evaluated based on reference to histopathology after surgery and/or clinical diagnostic criteria. Results Malignant stricture was finally diagnosed in 96 patients, and benign diseases in other 48. Brush cytology was positive in 78 of 96 patients with malignant stricture ( sensitivity 81.3% ), and negative in all patients without malignancy ( specificity 100. 0% ). Overall accuracy of diagnosis was 87.5%. Positive rate of malignancy by brush between 2007 and 2009 was 87.7% (50/57), while that between 2004 and 2006 was 71.8%, which were significantly different ( P ＜ 0. 05 ). The positive rate was not correlated with stricture location or tumor type. No major complications occurred, except for 4 moderate acute pancreatitis, 3 cholangiolitis and 2 biliary bleeding. Conclusion Brush cytology is of high sensitivity and specificity for malignant biliary stricture.Repeated brushing and manipulation can facilitate diagnosis yield.

Objective To clone the full-length of human Gill cDNA and construct the recombinant lentiviral expressing vector pLOX-Gfil for eukaryotic expression,providing a basis for further study on the biological functions of Gfil.Methods Total RNA was isolated from K562 cells,and the full-length Gfil cDNA was amplified by RT-PCR and then ligated with pGEM-T vector after retrieve and purification.The ligation product was transformed into competent cells DH5a.The positive recombinant clones were selected and identified by a complementation,restriction endonuclease digestion.The cloning vector and the lentiviral vector pLOX first digested with BarnH I were ligated and transformed.The enzyme and PCR analyses were performed to confirm the recombinant vector,and then DNA sequence analysis.Results A fragment of 1.2 kb was obtained by RT-PCR.The enzyme and PCR analyses revealed that the correct Gfil cDNA was cloned.The sequence of cloned cDNA was identical to the sequence deposited in GenBank (NM005263).Conclusion Gfil was cloned correctly and the recombinant lentiviral vector pLOX-Gfil for eukaryotic expression was constructed successfully.

Proteomic analysis of core needle biopsy (CNB) sample from patient populations is critical to our understanding of human disease,but has been hindered by its particular small size.Here,we present a method for the proteomic analysis of CNB sample based on the two dimensional electrophoresis.Proteins were extracted directly from 3 rat liver CNB specimens and a human prostate CNB sample.respectively.24 cm Immobiline DryStrip (pH 3-10NL) and 12.5% SDS-PAGE were introduced to separate the proteins.Interesting spots were analyzed by MALDI TOF/TOF mass spectrometry after tryptic digestion.With this method,consistent electrophoretic patterns of more than 2 500 protein spots were reproducibly obtained after silver staining,from rat liver CNB specimens.Qualitatively and quantitatively reproducible results also yield when the method was applied to a human prostate CNB sample.57 stochastically selected protein spots were analyzed by MALDI TOF/TOF moss spectrometry.and were identified with high confidence including faint ones.This simple and reproducible approach raises the opportunity of defining key molecular events of human disease pathologies.

Objective In order to evaluate the possible effects of myeloid-derived suppressor cells (MDSCs) on graft versus host disease (aGVHD) development and clinical outcomes,this study systematically detected the dynamic changes of MDSCs accumulation in patients during the first 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Peripheral blood was obtained from 30 patients and 10 healthy volunteers with heparin anticoagulant tubes for 6 mL.For patients,peripheral blood was collected during the first 100 days after allo-HSCT and MDSCs levels were detected by flow cytometry.For measuring the serum concentrations of IL-6,IL-10,IL-1β,TNF-α,Arg-1,HO-1 and iNOS,samples were analyzed using ELISA kits.Results Patients developing aGVHD were infused with significantly less number of MDSCs [(39.94 ± 8.383) 106/kg]than in those not developing aGVHD [(209.0 ± 57.68) 106/kg,P =0.002 6];Patients developing aGVHD Ⅰ-Ⅱ and patients without aGVHD received significantly greater number of MDSCs [(61.96 ± 13.67) 106/kg and (209.0 ± 57.68) 106/kg] than in those developing aGVHD Ⅲ-Ⅳ [(20.37 ±4.304) 106/kg,P =0.013 9].After allo-HSCT,the mean percentage of MDSCs increased markedly in patients developing aGVHD [(7.725 ± 1.460)％] as compared with those not developing aGVHD [(3.423± 1.044)％,P =0.021 3].The high MDSCs group (＞53.712 × 106/kg) showed more favorable clinical outcomes than in the low MDSCs group (≤53.712 × 106/kg).The 2-year overall survival rate as 100％ in high MDSCs group,and 50％ in low MDSCs group (P =0.001 3).The cumulative incidence of 2-year relapse was 6.250％ and 29.252％ in high MDSCs group and low MDSCs group respectively (P =0.112 3).The cumulative incidence of NRM was significantly lower in high MDSCs group (0％) than in low MDSCs group (49.519％,P=0.001 8).MDSCs frequencies significantly increased in patients developing aGVHD after allo-HSCT.After allo-HSCT,the concentrations of IL-6,IL-10,TNF-α,Arg-1,iNOS and HO-1 were significantly elevated in patients developing aGVHD.Conclusion The number of MDSCs when engraftment may be used as a predictor for the development and severity of aGVHD.MDSCs might be considered as a potential new approach to regulate transplant rejection and achieve long-term survival.

BACKGROUND:Glial scar and cavity formation fol owing spinal cord injury inhibits axonal entrance, so limited axonal regeneration, less secretion of neurotrophic factor and inhibitors in the microenvironment of axonal growth are considered as major impediments for impacting functional recovery of patients with spinal cord injury.
OBJECTIVE:To analyze literatures home and abroad related to the biological characters of astrocytes and glial scar hyperplasia after spinal cord injury, and to provide a theoretical basis for the mechanism underlying glial scar formation fol owing spinal cord injury.
METHODS:PubMed and Wanfang databases were retrieved using the keywords“astrocytes, reactive astrogliosis, glial scar, spinal cord injury”in English and Chinese, respectively. Final y 62 literatures were selected for overview.
RESULTS AND CONCLUSION:Currently, studies concerning the biological characters of astrocytes, reactive astrogliosis and glial scar formation fol owing spinal cord injury have achieved some progresses. Studies mainly focus on the sole impediment for spinal cord injury, and treatment also aims at inhibiting single factor, but interactions among factors have not been confimed. In addition, the regulatary mechanisms of specific intracel ular and extracel ular signal molecule in the astrocytes, and effective control and interference of glial scar formation fol owing spinal cord injury stil need in-depth study.