BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) under certain conditions can differentiate into a variety of connective tissue cells, also can be turned into neurons and glial cells. OBJECTIVE: To investigate biological characteristics of neural stem cells (NSCs) from adult bone marrow by immunomagnetic beads. DESIGN, TIME AND SETTING: Autologous bone marrow was selected as observational objects. Experiments were performed at the Department of Clinical Laboratory, Heze Municipal Hospital from March 2003 to June 2007. MATERIALS: Bone marrow was collected from inpatients with stroke, senile dementia, Parkinson's disease and cerebral ischemia at the Department of Neurology, Heze Municipal Hospital. METHODS: NSCs were harvested by immunomagnetic beads through isolating immunomagnetic beads from other cells under magnetic field following combination between specific antibody of immunomagnetic bead surface and NSC antigen in bone marrow. Purified NSCs were inoculated in Eagles MEM medium containing fetal bovine serum (FBS) of 0.20 volume fraction. Primary culture was performed in the medium in addition of basic fibroblast growth factors (bFGF) and epidermal growth factor (EGF). MAIN OUTCOME MEASURES: Immunohistochemistry and immunofluorescence were applied to identify the cells. Flow cytometry was used to measure DNA content. RESULTS: Under a phase contrast microscope, NSCs showed turgor vitalis, clear nucleus and nucleolus, enlarged nervous process and dense lattice. Immunohistochemistry and immunofluorescence demonstrated that neuron-specific enolase (NSE), neuroprotein, glial fibrillary acidic protein (GFAP) and microtubule-associated protein (MAP) were positive. Flow cytometry confirmed that DNA content was normal diploid, no induced mutation of the passaged cells. CONCLUSION: NSCs are harvested from adult bone marrow by combination between specific antibody of immunomagnetic bead surface and NSC antigen in bone marrow. NSCs have the characteristics of nerve cells in neurochemistry and morphology.

Objective To study the expression of basic fibroblast growth factor(bFGF),fibroblast growth factor receptor-2(FGFR-2)and heparanase(Hpa)in the tissues of gastric cancer and precancerous lesions,and to explore its significance in the carcinogenesis and malignant progression of stomach.Methods The expression of bFGF,FGFR-2 and Hpa were detected by immunohistochemistry in 145 cases of paraffin-embedded specimens from different gastric mucosa,including 30 cases of chronic superficial gastritis(CSG),29 of intestinal metaplasia(IM),31 of dysplasia(Dys)and 55 of gastric carcinoma(GC).Results The expression of bFGF and FGFR-2 in CSG group was significantly lower than that of the other three groups(P

Aim To explore the role of prenatal exposure to lipopolysaccharides(LPS)on aortic morphology in the 6-week offspring rats.Methods Twelve pregnant rats were randomly divided into three groups: control group,LPS group,L+P(pyrrolidinedithiocarbamate,PDTC)group.The rats were intraperitoneally administered vehicle,LPS(0.79 mg·kg-1),or LPS plus PDTC(100 mg·kg-1).LPS was given on the 8th,10th and 12th day,where as vehicle and PDTC were given daily from the 8th to the 14th day during gestation.Offspring body weight was measured at 6-week,and histopathological alteration of the thoracic aorta was observed by transmission electron microscopy,the thoracic aorta mRNA and protein expression of connexin(Cx)molecules including Cx37,Cx40,Cx43 and Cx45 in offspring rats were detected by real time PCR,Western blot and confocal laser-scanning microscope.Results Body weight at 6-week offspring rats was significantly higher in LPS group than in control group whatever male or female(P<0.01);and that in L+P group was significantly lower than in LPS group in male rats(P<0.05),but not in female rats.The LPS group thoracic aortas exhibited lesions,including impaired endothelial cells,and the vascular smooth muscle cells were arranged disorder and migration and proliferation;the number of gap junction was increased and longer than control group;the pathological changes were improved to some extent in L+P group.Cx43 mRNA and protein expression in LPS group was obviously lower than in control group(P<0.05),which could be partly reversed in L+P group(P<0.05).There was no significant difference in Cx37,Cx40 or Cx45 mRNA or protein expression among each group.The results of protein expression with confocal laser-scanning microscope was the same as those of Western blot.Conclusion Maternal LPS exposure during pregnancy leads to vascular changes in 6-week offspring rats,which might last until neonatal stage,and is closely correlated to hypertension in adult offsprings.

OBJECTIVE To investigate the causes of stomal recurrence after laryngectomy and the possibility of surgical management.METHODS The clinical data of 13 cases of stomal recurrence were analyzed retrospectively.RESULTS In 13 cases of stomal recurrence after laryngectomy,the survival period of 9 cases treated surgically was longer than that of 4 cases untreated or palliative treatment.CONCLUSION The overall prognosis of stomal recurrence after laryngectomy is poor.A proper surgical treatment in some cases will prolong the patient's lifetime obviously and improve his quality of life.

Objective To explore preliminarily the role of the E2 subunit of pymvate dehydrogenase (PDC-E2) modified by xenobiotics (e.g.2-octynic acid,2-OA) in the pathogenesis of primary biliary cirrhosis (PBC).Methods Patients of PBC (102 cases),primary sclerosing cholangitis (PSC,34 cases) and healthy controls (HC,50 cases) were selected.The anti-PDC-E2,anti-2-OA and anti-lipoic acid (LA) antibody in the peripheral blood of the 3 groups were tested by enzyme linked immunosorbent assay (ELISA).By inhibitive ELISA (iELISA),30 of the 102 PBC patients with anti-PDC-E2 antibody but without anti-2-OA antibody were selected to detect whether there was any new epitope on the PEC-E2 conjugated with 2-OA.The chi-square test and Fisher exact test were taken to analyze the enumeration data.The two-tailed unpaired t test with Welch's correction was used to compare the measurement data.Spearman rank correlation analysis was also employed for proper test.Results The positive rate of anti-PDC-E2,anti-LA and anti-2-OA antibody in PBC patients was 94.1％(96/102),73.5％(73/102) and 53.9％(55/102) respectively,all of which were statistically significantly higher than those in healthy controls group but were of no significant difference between PSC and healthy controls group.There was no significant relevance between the levels of Anti-LA and anti-2-OA antibody in the PBC group (r=-0.065,P=0.520).The iELISA results showed that the antibody,which only identified the epitopes on 2-OA-PDC-E2 induced by the 2-OA conjugation with PDC-E2,existed in 40％(12/30) of the PBC patients,and more interestingly,this antibody was predominantly appeared in PBC patients at their early clinical stage.Conclusion There are anti-LA antibody and anti-2-OA antibody in PBC patients,which have shown no significant association with each other.It is very likely that new antigenic conformational epitopes on PDC-E2 modified by 2-OA would emerge,which might led to the immune response in the individuals who are susceptible to PBC,and thus contribute for the breaking of PDC-E2 immune tolerance,and PBC occurrence finally.

Objective To determine the levels and significance of Krebs von den lungen-6(KL-6), pulmonary surfactant protein A (SP-A), SP-D and interleukin (IL)-6 in patients with connective tissue disease interstitial lung disease (CTD-ILD). Methods The serum KL-6, SP-A, SP-D and IL-6 in all subjects were detected and the imaging and pulmonary function were recorded t test, χ2 test, non-parametric test, ANOVA and correlation analysis were used for data analysis. Results ① The levels of serum KL-6, SP-A, SP-D, IL-6 in the CTD-ILD group [551.4 (428.2, 883.5) U/ml, 938.4(435.2, 2324.7) pg/ml, 90.7 (80.7, 100.3) ng/ml and 30.4 (22.9, 41.7) pg/ml; P all<0.05] was significantly higher than that in the CTD group [192.9 (139.2, 266.2) U/ml; 458.0 (372.6, 529.0) pg/ml; 80.0 (71.2, 98.3) ng/ml; 18.6 (4.9, 31.0) pg/ml, Z=-5.383, -3.76, -2.123,-3.903, P all <0.05]; and higher than healthy controls (n=30) [183.2(141.9, 216.6) U/ml; 229.0(162.0, 248.0) pg/ml;50.8(26.1, 96.4) ng/ml;7.1(3.7, 8.7) pg/ml, Z=-5.801,-8.13, 2.272, 3.266;P all<0.05].②The levels of KL-6 in pulmonary HRCT for active ILD group was significantly higher than the non-active ILD group [998.5 (640.3, 1293.3) U/ml vs 565.0(434.0, 799.5) U/ml, Z=2.182, P=0.023], there was no statistical difference in the levels of SP-A, SP-D, IL-6 between the 2 groups. ③ Spearman correlation analysis showed that KL-6 was negatively correlated with forced vital capacity (FVC%);SP-D, IL-6 and diffusing capacity of carbon monoxide (DLCO %). ④ Logistic multiple regression analysis showed that KL-6 [OR=1.017, P=0.002, 95%CI (1.006, 1.028)], SP-A [OR=1.023, P=0.009, 95%CI (1.006, 1.041)], SP-D [OR=1.175, P=0.009, 95%CI (1.075, 1.264)], IL-6[OR=1.213, P=0.001, 95%CI(1.088, 1.354)] were the risk factors for ILD. Conclusion Serum KL-6, SP-A, SP-D and IL-6 are significantly increased and correlate with CTD-ILD. KL-6 is related to the pulmonary inflammatory disease and vital capacity, while SP-D and IL-6 are related to diffusion function.

Objective To characterize the clinical characteristics of lupus mesenteric vasculitis (LMV). Methods Analyzing the clinical, laboratory and treatment data of LMV patients hospitalized from 2002. 1.1 to 2007. 12. 31 retrospectively. Results (1) The three common manifestations were abdominal pain, diarrhea and vomit with the prevalence rate of 77%, 70% and 67% respectively. (2)The majority of LMV cases were active vital organ (28/30), kidney (24/30) and hematological system (18/30) were the main organs of involvement. Ten patients had hydroureteronephrosis, and 8 patients had intestinal pseudo-obstruction at the same time. (3) Systemic lupus erythematosus disease activity index (SLEDAI) score was ≥10 in 80% (24/30) of patients. The progression of LMV was accompanied with new-onset ieucopenia or worsening leucopenia or hypocomplementemia in 10 cases. (4) Blood antinuclear antibodies were positive in 27 patients detected, and anti-SSA antibody was positive in 15 (56%), anti-U1RNP antibody was positive in 14 (52%). (5) Fourteen cases had bowel wall thickening with target sign or mesenteric vessels with palisade or comb sign in contrast CT scan of abdomen. (6)Twenty-seven cases were treated with orally or intravenous medium to high dose steroid therapy and recovered from LMV. Conclusions (1) Abdominal pain, diarrhea and vomit were frequent manifestations of LMV patients. (2) LMV was one of the serious complications of systemic lupus erythematosus(SLE), and usually accompanied by active SLE in other organs. (3) A drop in the white blood cell count or complement C3 titer might be correlate with the occurrence of LMV. It needs to further investigate the relationship between LMV and the high positive rate of anti-SSA and anti-U1RNP antibody. (4) LMV patients responded well to intravenous high dose methylprednisolone.

Objective To evaluate the therapeutic effect of endoscopic management of plastic stents of post-liver transplantation anastomotic biliary stricture. Methods From January 2010 to October 2015, clinical data of patients with post-liver transplantation anastomotic biliary stricture and received endoscopic retrograde cholangiopancreatog﹣raphy and plastic stents management was collected. The technical success rate, ERCP-related complications, clinical remission rate and long-term complications were main outcome measurements to compare the efficacy and safety of different number of stents in managing post-liver transplantation anastomotic biliary stricture. Results Among the 18 patients (0.5 ~ 60.0 months) with post-liver transplantation ABSs, seven patients received less plastic stents treat﹣ment (< 3 stents), nine patients with persistent anastomotic or recurrent stricture received multiple plastic stents treatment (≥ 3 stents), two patients received multiple plastic stents treatment once suffered with post-liver trans﹣plantation ABSs. The endoscopic technical success rate was seventy-six over eighty (95.0 %). Among the seven pa﹣tients received less plastic stents treatment, one loss to follow-up, two were still under treatment, one died of acute hepatic failure, one died of septic stock, one combined with biliary fistula resulted in treatment failure, one achieved clinical remission, the clinical remission rate was one third (33.3 %). Among the eleven patients received multiple plastic stents treatment, two loss to follow-up, one was still under treatment, two received surgery because of failed treatment, six achieved clinical remission, the clinical remission rate was 75.0 % (6/8). The average diameters and stent durations of management of 1 stent, 2 stents, 3 stents, 4 stents, 5 stents, 7 stents were 8.5 F, 17.0 F, 24.0 F, 28.0 F, 36.0 F, 50.0 F. Among the six early postoperative complications, five cases occurred in less stent manage﹣ment and one occurred in MPSs management, the early postoperative complication rate was 7.5 %(6/80). No severe ERCP-related complications and procedure-related deaths. Conclusions Endoscopic management of plastic stents is safe and effective for post-liver transplantation ABSs. Providing larger biliary support, the multiple plastic stents treatment was superior to less plastic stents treatment in view of clinical remission rate, especially for refractory one. Multiple plastic stents did not increase the incidence of complications, it could be used as the first-line treatment of post-liver transplantation duct-to-duct biliary anastomosis for its safety and effectivity.

Objective To study the diagnostic value of endoscopic retrograde cholangiopancreatography (ERCP) combined with intraductal ultrasonography (IDUS) for bile duct stricture diseases.Methods The results of endoscopic ultrasonography and endoscopic retrograde cholangiopancreatography,intraductal uhrasonography,bile duct brushing cytology,the liquid-based cytology and the histopathological examination were analyzed retrospectively.The final diagnosis was made based on clinical data,histopathology and follow-up results(≥4 months).Results Twenty-one patients were diagnosed as having malignant biliary diseases,including 9 biliary tract carcinoma,4 duodenal papilla carcinoma,4 pancreatic cancer infiltrated common bile duct,4 cancer of the liver infiltrated common bile duct ; 15 were diagnosed as having benign biliary diseases,including 9 bile duct stones,4 liver fluke disease,lbile duct inflammatory stenosis,1 bile duct stricture caused by external compression.The accuracy rate of the EUS,ERCP,IDUS and ERCP + IDUS in the differential diagnosis of the bile duct stricture disease were 77.8％,88.9％,91.7％ and 94.4％,respectively.The accuracy rates of IDUS,ERCP,ERCP combined with IDUS were significantly higher than that of EUS (P ＜ 0.05).The sensitivity,specificity,positive predictive value and negative predictive value of ERCP combined with IDUS were 95.2％,93.3％,95.2％ and 93.3％ respectively which were higher than those of EUS,ERCP and IDUS.After the bile duct brushing cytology and the liquid-based cytology or histopathological examination,19 patients were diagnosed as having malignant biliary diseases,17 were benign biliary diseases.The sensitivity,specificity and accuracy of the procedures for malignant bile duct stricture disease were 90.5％,100.0％,94.4％ respectively.Conclusion ERCP combined with IDUS can improve the diagnostic accuracy.The diagnostic positive rate will be higher with the help of ERCP,IDUS and targeted brush.

Objective To evaluate the diagnostic value of the cell block (CB) with immunostaining method by endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) biopsy for pancreatic lesions.Methods A total of 72 patients with pancreatic lesions underwent EUS-FNA at the First Affiliated Hospital of Guangxi Medical University from March 2012 to June 2013.The EUS-FNA samples of all patients were processed by conventional smear cytology,liquid-based cytology (LBC) and the cell block with immunostaining.Results There were 61 pancreatic patients who were finally diagnosed as having pancreatic tumors,including 55 cases of pancreatic cancer,2 pancreatic solid pseudopapillary tumor,4 pancreatic endocrine tumors (PETs),and 11 benign lesions:4 chronic pancreatitis,2 pancreatic tuberculosis,4 pancreatitis and 1 pancreatic mucinous cystadenoma.The diagnostic sensitivity of conventional smear cytology,liquid-based cytology and cell block with immuno-staining method were 68.9％ (42/61),75.4％ (46/61) and 90.2％ (55/61),respectively.The diagnostic specificity of three methods were all 100.0％.The diagnostic accuracy rates were 73.6％ (53/72),79.2％ (57/72) and 91.7％ (66/72),respectively.The diagnostic accuracy rate of the cell block with immunostaining was higher than those of conventional smear cytology (P ＜ 0.05) and the liquid-based cytology (P ＜ 0.05).Conclusion Endoscopic ultrasound-guided fine-needle aspiration is a safe and effective method with high sensitivity and specificity in the diagnosis of pancreatic lesions.Cell block method combining immunohistochemistry helps to increase the diagnosis and histological diagnosis of pancreatic lesions.The cell block has a greater clinical value in the diagnosis of pancreatic lesions.