AIM:To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats .METHODS: Male SD rats (n=48) were randomly divided into sham group , ischemia/reperfusion (I/R) group, RNA interference group and nega-tive interference group .The rat middle cerebral artery was blocked to establish focal cerebral I /R model ( ischemia for 1 h and reperfusion for 7 d).Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain.The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB 1/GFAP and Western blotting .The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labe-ling of BrdU/nestin.RESULTS: The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05).RNA interference effectively inhibited the HMGB1 expression (P<0.05).Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05).The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05).CONCLUSION:Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB 1 protein level .

Objective To investigate the effect of CXCL16 on the development of murine collageninduced arthritis (CIA). Methods CXCL16 mRNA of the involved synovium and serum CXCL16 protein were determined respectively by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA) in murine collagen-induced arthritis. The proliferation of lymphocytes from murine spleen and the level of RANKL mRNA, stimulated by CXCL16 at different concentrations (0,100, 200, 400, 800 ng/ml), was detected respectively by CCK-8 and RT-PCR, then the level of IL-2 and IFN-γ in culture supernatant was detected by ELISA. Comparisons between groups were tested by t test and one-way ANOVA analysis. Results The serum CXCL16 [(127± 10) vs (72±8) pg/ml, P＜0.05] and synovial CXCL16 mRNA (0.214±0.007 vs 0.375±0.009, P＜0.01) in CIA were all significantly higher than those in normal controls. The proliferation of CXCL16 (200, 400, 800 ng/ml) in CIA mouse lymphocytes, was significantly higher than that of CXCL16 (0 ng/ml) (0.51±0.06, 0.56±0.05, 0.55±0.04, (0.41±0.04, P＜0.05). And CXCL16 on the CIA stimulated lymphocyte proliferation was significantly higher than controls on normal lymphocytes (P＜0.05). Compared with blank control group, the expression of IL-2, IFN-γ and RANKL mRNA of CIA CXCL16 (400, 800 ng/ml) groups was higher significantly (P＜0.01). Conclusion CXCL16 plays an important role in the development of murine CIA by activating lymphocytes.

ObjectiveTo establish the reference value of inhibin-A in the serum of pregnant women with gestational age from 15 to 20+6 weeks in Guangdong province,and assess the efficiency of inhibin-A and quadruple test in Down syndrome screening.Methods A total of 2802 singleton pregnancies receiving triple test screening in Guangzhou Women and Children's Medical Center from March 2008 to December 2010 were included in this study.Inhibin-A was measured by automatic enzyme-linked immunosorbent chemiluminescence assay. The concentration of inhibin-A was expressed as multiples of the median (MoM),and adjusted with maternal weight and gestational age.Parameters of SURUSS were used to recalculate the risk of Down syndrome.The efficacy of single marker and combination were evaluated by receiver operating characteristic curve and the area under the curve. Results(1) In normal singleton pregnancies,the median concentration of inhibin-A was 286.60,267.10,249.10,243.40,242.30 and 256.60 pg/ml respectively for each week of gestational age from 15 to 20+6 weeks.The distribution of inhibin-A in each gestational week was relatively stable.The mean concentration [(852.83±370.04) pg/ml] and MoM (2.82) of inhibin-A in twelve pregnant mothers with Down syndrome fetuses were significantly higher than those without [(293.28±149.46) pg/ml (t=5.37,P＜0.05) and 1,respectively].(2) The detection rate was 83.3％ (10/12) by using the quadruple test including free human chorionic gonadotropin-β,alphafetoprotein,unconjugated estriol and inhibin-A at false positive rate of 5.8％; while when the detection rate of triple test including alpha-fetoprotein,free human chorionic gonadotropin-β and unconjugated estriol was 83.3％,the false positive rate was 7.7％.When the false positive rate was set to 5.0％,the area under the curve of inhibin-A,alpha-fetoprotein,free human chorionic gonadotropin-β and unconjugated estriol was 63.7％,20.5％,46.1％ and 4.8％,respectively,and the relative area under the curve of routine triple test and quadruple test was 45.5％ and 63.1％,respectively.ConclusionsInhibin-A is suggested to be the most effective marker used for secondtrimester screening,which could be used for second trimester Down syndrome screening in Chinese population combined with existing three markers.

Objective To diagnose a rare variant Hb Shaare Zedek in a Chinese family,and describe its clinical feature.Methods Blood samples of seven members of three generation were collected.Standard blood routine analysis,hemoglobin electrophoresis and blood gas analysis were used for phenotype analysis.Gap-PCR and reverse dot blot analysis were used to detect common thalassemia mutations.DNA sequence analysis of the human α and β globin genes were used to identify the mutation site of these samples.Results The result of blood routine analysis was normal in the proband,but a abnormal hemoglobin band (22.4％,capillary assay) was found by hemoglobin electrophoresis.In the blood gas analysis,the partial pressure of oxygen and blood oxygen saturation were 72.0 mm Hg ( 1 mm Hg =0.133 kPa) and 93.0％ in the proband.The heterozygous mutations in αl globin gene at codon 56 ( AAG ＞ GAG)which leaded Glu substitution to Lys were identified in the proband.Other family members who carried the same mutation showed similar phenotype,with abnormal hemoglobin band ( 22.4％ - 23.9％,capillary assay),low partial pressure of oxygen (59.0 - 72.0 mm Hg) and blood oxygen saturation (91.0％ -93.0％).Conclusions The heterozygote of Hb Shaare Zedek leads to slight symptoms with abnormal hemoglobin band,decreased partial pressure of oxygen and blood oxygen saturation.Discovery of this mutation enriches the abnormal hemoglobin spetrum of Chinese people,and it is useful for the clinical diagnosis and genetic counseling of hemoglobinopathies.

Objective:To investigate the aberrant methylation of CpG island in 5′promoter region of p16 gene in the pancreatic juice and its value in diagnosis of patients with pancreatic cancer.Methods:Pure pancreatic juice(PPJ)was collected from the pancreatic duct by a nasopancreatic tube put under endoscopic retrograde cholangiopancreatography(ERCP). Cytological examination was performed by H-E staining in pure pancreatic juice.Aberrant p16 methylation was detected using the methylation specific PCR(MSP)in the PPJ.Results:The sensitivity,specificity,positive predictive value,negative predictive value and accuracy cytological examination in diagnosis of pancreatic cancer were 40%,100%,100%,45.4% and 60.0%,respectively.The DNA sequences were successfully extracted from the PPJ of 30 patients with pancreatic diseases and were subjected to MSP.Seven(35%)of the 20 cases with pancreatic cancer showed aberrant methylation of p16 gene.No aberrant methylation was detected in the pancreatic juice samples of patients with chronic pancreatitis and mucinous cystoadenocarcinoma of pancreas.When cytological examination combined with p16 methylation detection,the sensitivity, specificity,positive predictive value,negative predictive value and accuracy for diagnosis of pancreatic cancer were 55%,100%, 100%,52.6% and 70%,respectively.Conclusion:Pancreatic juice collected by nasopancreatic drainage during ERCP can be used for molecular analysis.Detection of aberrant methylation of p16 gene in pancreatic juice combined with cytological examination is a better method for diagnosis of pancreatic cancer.

OBJECTIVETo investigate the renal protective effects of sulodexide and its anti-oxidative stress mechanism in diabetic rats.

METHODThirty male SD rats were randomized into 3 equal groups, namely the control group, diabetic group, and sulodexide treatment group. Twelve weeks after establishment of rat diabetic models and administration of sulodexide, the rats were sacrificed for measurement of the urine volume, body mass, kidney mass/body weight ratio, plasma glucose, and glycosylated hemoglobin (HbA1c). Malondialdehyde (MDA) levels and superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) activities in the renal tissue or serum were tested. Electron microscopy was performed to observe the pathological changes in the kidneys.

RESULTSThe urine volume, renal mass/body mass ratio, serum glucose, HbA1C, and serum and renal MDA levels all significantly increased in the diabetic rats in comparison with the normal controls (P<0.05). But the body weight and activities of SOD, CAT, and GSH-PX in the renal tissue in the normal control group were significantly higher than those in the diabetic and sulodexide group. After 12 weeks of sulodexide treatment, SOD, CAT, and GSH-PX activities in the renal tissue of rats were significantly increased in comparison with those in the diabetic rats (P<0.05). Electron microscopy showed obvious irregular thickening of the glomerular capillary basement membrane in the diabetic group with vacuolization in the mitochondria in the epithelial cells, and such pathological changes were significantly alleviated in the sulodexide treatment group.

CONCLUSIONSSulodexide can effectively lower the urinary albumin excretion rate, improve the ultrastructural renal pathologies and prevent glomerular basement membrane thickening in diabetic rats, probably in association with the reduction of the MDA levels and enhancement of SOD, CAT, and GSH-PX activities.

Animals ; Antioxidants ; pharmacology ; therapeutic use ; Body Weight ; drug effects ; Catalase ; metabolism ; Diabetes Mellitus ; drug therapy ; metabolism ; pathology ; Glutathione Peroxidase ; metabolism ; Glycosaminoglycans ; pharmacology ; therapeutic use ; Kidney ; drug effects ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Organ Size ; drug effects ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the effect of combining the injection of beta-sheet breaker H102 with human umbilical cord mesenchymal stem cell (hUCMSC) on APP transgenic mice behavior, P-tau, apoptosis and the expression of relevant enzymes in the brain.

METHODSAPP transgenic mice were randomly divided into model group, hUCMSC group, H102 group, H102 with hUCMSC group and a group of C57BL/6J mice with the same age and background was set as normal. After two weeks and four weeks, the ability of spatial reference memory was tested by Morris Water Maze. After four weeks, immunohistochemical stain and Western blot were done to detect the content of Bad, Bax, Bcl-2, Bcl-xl, P-tau, GSK-3beta, PP-2A and PP-1 in mice brain.

RESULTSThe ability of memory of hUCMSC in 2 weeks group was slightly improved than that in the model group. hUCMSC in four weeks group, H102 group and H102 with hUCMSC group significantly improved the ability of and memory, and reduced the phosphorylation of tau and brain cell's apoptosis of the Alzheimer disease (AD) mice.

CONCLUSIONBeta-sheet breaker H102 together with transplanting hUCMSC is an effective therapeutic strategy for AD.

Alzheimer Disease ; therapy ; Amyloid beta-Protein Precursor ; genetics ; Animals ; Disease Models, Animal ; Humans ; Maze Learning ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Peptides ; therapeutic use ; Umbilical Cord ; cytology

OBJECTIVETo improve the ability of recognizing, diagnosing and treatment for the femoral shaft fractures combined with ipsilateral occult femoral neck fractures, and reduce the rate of loss-diagnosis.

METHODSBy retrospective study of 50 patients who sustained ipsilateral femoral shaft and neck fractures from March 1998 to October 2003, 9 cases were femoral shaft fractures combined ipsilateral femoral neck fractures. The neck fractures were diagnosed separately before, during and after operation. Among the 9 cases, 5 cases treated with reconstructive intramedullary nail to fix both shaft and neck fractures; 3 cases were treated with retrograted intramedullary nail to fix shaft fractures and with canulated screws to fix neck fractures; another one was sustained two operations, after the fixation of shaft by intramedullary nail, the neck fracture was found, the neck fracture was fixed with canulated screws anterior and posterior of the nail.

RESULTSNine cases were followed up for average 20 months. All femoral shaft fractures were united during 6 months; and all neck fractures were united during 3 months.

CONCLUSIONSFemoral shaft fractures combined with ipsilateral occult femoral neck fractures have a high rate of loss-diagnosis during the early stage after injure, the orthopaedic surgeons should consider the femoral neck fracture by analyzing the mechanism of high energy injury patient with femoral shaft fracture. The occult femoral neck fractures can be diagnosed by CT scan before operation. During and after the operation of fixing the femoral shaft fractures, the femoral neck should be observed intensively to identify the exist of fractures.

Adult ; Femoral Fractures ; complications ; diagnostic imaging ; surgery ; Femoral Neck Fractures ; complications ; diagnostic imaging ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; methods ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed

OBJECTIVETo investigate the rate of systemic inflammatory response syndrome (SIRS) among patients treated by intramedullary nail (IMN) for femoral shaft fracture right after the operation, the effect of femoral nailing on SIRS and sought to differentiate any differences between reamed and unreamed IMN.

METHODSThree hundred and twenty-four patients presenting with acute femoral fractured from April 1997 to April 2005, divided into 2 group depending on ISS. Group 1 had 179 patients (ISS < 16, AIS < 2) including 64 reamed cases and 115 unreamed cases; group 2 had 145 patients (ISS >or= 16, AIS >or= 2) including 51 reamed patients and 94 unreamed. All patients were treated by close reduction and intramedullary nail for fixation. The heart rate, breath rate, temperatures were recorded every 6 h before and after operation. If any of the life sign was above normal, then venous blood samples were taken at once to make sure whether SIRS occurred or not. All data were crosstabs analysis by SPSS 10.0.

RESULTSOne hundred and twenty-four patients suffered from SIRS postoperation. In group 1, 24 reamed and 37 unreamed, in group 2, 27 reamed and 36 unreamed. Among those SIRS, 119 cases healed evenly and 5 patients developed further complication. Between reamed and unreamed patients both in group 1 and group 2, there was no significant difference found. The Chi-Square value were 0.159 and 2.885 respectively, total value was 3.467, which P was 0.325, 0.089 and 0.471, all greater than 0.05.

CONCLUSIONSAlthough SIRS will result from IMN, there is no significant different effect on the rate of SIRS found between reamed and unreamed nail in both group.

Adult ; Bone Nails ; adverse effects ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Intramedullary ; adverse effects ; methods ; Fractures, Closed ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Complications ; etiology ; Retrospective Studies ; Systemic Inflammatory Response Syndrome ; etiology ; Time Factors

Objective To screen the active compounds targeting lung cancer stem cells (LCSCs) from the sponge Aaptos aaptos.Methods The A549-Nanog-GFP model of LCSCs was constructed.Western blot and immunofluorescence were used to examine the expression of pluripotency markers in screening model.The established LCSCs model was used to screen 8 fractions of Aaptos aaptos dichloromethane extract.The active fraction was separated by various chromatographic methods.CCK8 assay was used to screen the compounds for anti-LCSCs activity in vitro.Results LCSCs model with high expression of CD44 and ALDH1A1 protein was successfully constructed.The fraction D6 showed significant inhibitory activity in LCSCs(P<0.01).Four aaptamine alkaloids were isolated from this fraction.Compound AP-1 has good activity against LCSCs(P<0.01)with IC50 value(3.84±0.12) μmol/L.Conclusion AP-1 isolated from the spongeAaptos aaptos exhibited significant activity against LCSCs.