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.

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 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

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 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 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.

ObjectiveTo investigate the clinical characteristics and predisposing factors of systemic lupus erythematosus(SLE) with sepsis.MethodsTwenty-one SLE patients with sepsis admitted to our hospital between 2005-2010 were reviewed in this study.The other 21 inpatients with active SLE in our hospital in the same period were randomly selected as controls.Clinical and laboratory documents of these patients were comparatively analyzed.Results The peak body temperature [ (39.4±0.6) vs (37.2±0.4) ℃,t=13.403,P=0.000],the hyperpyrexia (T≥39 ℃) incidence (71％ vs 5％,X2=19.788,P=0.000),the white blood cell (WBC) counts[(10.2±4.6) vs(6.2±2.5)×109/L,t=3.469,P=0.001)] and neutrophils in the peripheral blood [(8.3±4.5) vs(4.5±2.1)×109/L,t=3.559,P=0.001 ],the C-reactive protein(CRP) level [ (74±59) vs (5±4) mg/L,t=5.398,P=0.000 ] and lactate dehydrogenase (LDH) level[ (444±343) vs ( 225±144) U/L,t=5.398,P=0.000] in the sepsis group were significantly higher than those in the control group.It was noticeable that CRP in the sepsis group was 15 to 20 times higher than that in the control group.The level of serum albumin[ (29±9) vs(35±7) g/L,t=2.688,P=0.011 ],the maintenance dosage of hydroxychloroquine [(0.11±0.08) vs(0.17±0.09) g/d,t=2.331,P=0.025],the frequency of autoantibodies against SSA (38％ vs 71％,X2=4.709,P=0.03) or SSB(0 vs 43％,X2=11.455,P=0.001) in the sepsis group were significantly lower than those in the control group.Correlation analysis showed that,in the sepsis group,the SLE disease activity index (SLEDAI) had significant positive association with SLE duration (r=0.514,P=0.017),the WBC count (r=0.552,P=0.010) and neutrophils count(r=0.545,P=0.011 ),respectively.The neutrophil count correlated positively with the LDH(r=0.482,P=0.032) and CRP(r=0.606,P=0.022).These correlations were not statistically significant in the control group.ConclusionSepsis should be considered when SLE patients have hyperpyrexia,high levels of WBC and neutrophils,markedly elevated LDH and CRP level,especially when the CRP increases ten times higher than the normal limit.SLE activity and sepsis might affect each other,and this may be more evident in patients with longer disease duration.Hypoalbuminemia and negative autoantibody to SSA or SSB are likely to be the risk factors for SLE to develop sepsis while hydroxcyhloroquine may be protective against sepsis.

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.

Objective To test the level of cell factor interleukin (IL)-21,CXCL13 in the plasma of patients with rheumatoid arthritis (RA),and to analyze the relationship between Follicular helper T cells(Tfh)and clinic features and discuss the possible immunological pathogenesis of RA.Methods The Tfh cells were obtained from patients and healthy controls (NC) and detected by Flow cytometery.While the levels of IL-21,CXCL13 in patients and NC were measured by ELISA tests.Those analysis were performed by student's t-test,one-way ANOVA,SNK-q test,Chi-square test,Spearman's correlation and multiple linear regression.Results The expression of CD4+CXCR5+ICOS+ cells (Tfh) in PBMCs of RA was significantly higher than normal controls (3.0±1.2 vs 1.1±0.4,P＜0.01).Meanwhile,the three RA groups of patients were divided to low,moderate and high disease activity groups,and the results showed that the expression of Tfh were increased accordingly (1.8±0.7,2.5±0.6,4.0±1.2).The expression of Tfh in the three groups were all significantly higher than that of controls (P＜0.01).There was a positive correlation between Tfh and DAS28,ESR,CRP,TJC,and bone erosion,RF and anti-CCP respectively.The expression of Tfh in those patients who had bone destruction was higher than those with no or mild bone destructions (2.7±1.1vs 3.4±1.3).The expression of Tfh in patients with un-treated RA patients,when compared to those RA patients who were treated appropriately and those who were not treated appropriately,was decreased significantly.The expression of Tfh in appropriately treated RA patients was lower than that without appropriately treatment.The level of IL-21,CXCL13 was decreased in patients with RA in the order of high,moderate,low disease activity and NC.Conclusion The expression of Tfh and the levels of IL-21,CXCL13 are increased significantly,and are closely related to disease activity and bone ersions.The expression of Tfh is decreased after relevant treatment.These results indicate that the abnormality of Tfh may play an important role in the pathogenesis of RA.

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.