Objective To explore the safety of the mobilization of peripheral blood stem cells(PBSC)by granulocyte colony-stimulating factor(G-CSF)in elderly donors.Methods 28 peripheral arteriosclerotic occlusive disease(PAOD)elderly patients(aged≥60 years),and 29 healthy sibling young/adult donors(aged＜60 years)for peripheral allogenic stem cell transplantation were included.Blood samples were collected immediately before starting G-CSF and prior to PBSC collection to analyze the following parameters:the WBC counts,fibrinogen(FIB),D-dimer (D-D),thrombin antithrombin complex(TAT),antithrombin(AT)and yon Willebrand factor antigen(vWF:Ag). Results It had a very significant increase in D-D and vWF:Ag and a very significant decrease of AT(P＜0.01),af- ter mobilization by G-CSF,and a increase in FIB and TAT were also observed(P＜0.05,P＜0.01)in elderly group.In the young/aduh group,the increase in FIB was significant(P＜0.05).The elevating extent of D-D and TAT after G-CSF administration was significantly higher in elderly group than that in young/adult group(P＜0.05).Compared to young/adult group,there was a significant increase in thrombotic events and cerebrovascular ac- cident(P＜0.05).Conclusion In PBCS donorsreceiving G-CSF it reveals activation of both coagulation and en- dothelial cells and inhibition of anticoagulant system that can favor the developing of thrombotic events,which is more remarkable in elderly donors.Therefore a careful monitoring for coagulation system should be considered in those elderly cases.

AIMTo report the clinical experience during collecting sperm samples in the Fragile X syndrome (FXS) male patients.

METHODSTwo different polymerase chain reaction (PCR) based methods were used for the molecular diagnosis of FXS. Sperm collection was done mostly according to the laboratory manual of the World Health Organization.

RESULTSWe failed to collect sperm samples from five Fragile X subjects aged 18-60 years as a result of an unexpected erectile dysfunction (ED). Multiple examinations of the same subject at different times, and of different subjects from different provinces examined by different physicians, showed the same result consistently in all the five subjects.

CONCLUSIONErectile reflex is an instinctive response in all healthy males. The absence of erection can be caused by hormonal, physical or neuronal malfunction. As hormonal profiles were reported to be generally normal in Fragile X men, we propose that an unknown physical factor or the neuronal circuit, or both, underlying the erection is compromised. The finding of this symptom in Fragile X patients may help better understand the clinical spectrum and pathogeneses of the disease.

Adolescent ; Adult ; Base Sequence ; DNA Primers ; Erectile Dysfunction ; Female ; Fragile X Syndrome ; physiopathology ; Humans ; Male ; Middle Aged ; Pedigree ; Polymerase Chain Reaction ; methods ; Spermatogenesis ; genetics

OBJECTIVETo observe the expression of dynamin-1 and phosphor-dynamin-1 in the hippocampus of children and rats with mesial temporal lobe epilepsy (MTLE) and to investigate the roles of dynamin-1 and phosphor-dynamin-1 in the development of MTLE.

METHODSMale Sprague-Dawley rats (aged 25 days) were randomly divided into acute control (AC), acute seizure (AS), latent control (LC), latent seizure (LS), chronic control (CC) and chronic spontaneous seizure (CS) groups. Lithium chloride-pilocarpine was used to induce a rat model of MTLE. The hippocampus samples of 5 children with a pathologically confirmed hippocampal sclerosis who received surgical operation were collected as a human model (HM) group, and the hippocampus samples of 4 dead children (without organic lesion of the hippocampus) were collected by autopsy as a human control (HC) group. The expression of dynamin-1 and phosphor-dynamin-1 in the hippocampus of children and rats with MTLE was measured by Western blot and immunohistochemistry.

RESULTSThe Western blot showed that the expression of phosphor-dynamin-1 was significantly lower in the AS and CS groups than in the corresponding control groups (AC and CC groups) (P<0.05). The expression of phosphor-dynamin-1 was significantly lower in the HM group than in the HC group (P<0.05). There were no significant differences in the expression of dynamin-1 among the AS, LS and CS groups and between the HM and HC groups (P>0.05). The immunohistochemical results showed that phosphor-dynamin-1 was highly expressed in the cytoplasm of hippocampal neurons of AC, CC and HC groups, but its expression was significantly reduced in the AS, CS and HM groups (P<0.05).

CONCLUSIONSThe expression of phosphor-dynamin-1, not dynamin-1, is downregulated in the hippocampus of children and rats with MTLE during seizures, which suggests that the phosphorylation/dephosphorylation of dynamin-1 may be involved in the development of MTLE.

Animals ; Blotting, Western ; Child ; Dynamin I ; analysis ; metabolism ; Epilepsy, Temporal Lobe ; metabolism ; Female ; Hippocampus ; chemistry ; metabolism ; Humans ; Immunohistochemistry ; Male ; Phosphorylation ; Rats ; Rats, Sprague-Dawley

Objective To explore the roles of frequency domain OCT in measuring the retinal thickness around the optic disc and optic disc parameters in early glaucoma diagnosis.Methods The optic disc parameters and retinal nerve fiber layer (RNFL) thickness in the 40 healthy volunteers (control group) and 85 cases of primary open angle glaucoma (POAG),including 36 patients as the early glaucoma subgroup and 49 patients as the glaucoma evolutum subgroup,were measured by frequency domain OCT.Then,the correlation analysis of RNFL thickness,optic disc parameters and the mean deviation (MD) of visual field in each group was performed,and the area under the curve was used to evaluate the diagnostic efficiency of RNFL thickness around the optic disc and optic disc parameters in the diagnosis of glaucoma.Results The RNFL thickness and the complete cycle mean RNFL thickness in the temporal,upper,nasal and inferior quadrant in the glaucoma patients were significantly lower than those in the controls (all P ＜ 0.05),and the above indexes in the glaucoma evolutum subgroup were significantly decreased compared with those in the early glaucoma subgroup (all P ＜ 0.05).There were statistically significant differences in the optic disc parameters between the groups except the optic disc area (all P ＜ 0.05).Pearson correlation analysis showed the RNFL thickness and the complete cycle mean RNFL thickness in the temporal,upper and inferior quadrant were negatively correlated with the MD in the glaucoma patients (all P ＜ 0.05),and the parameter of optic cup volume and cup/disc area ratio were positively correlated with the MD (both P ＜ 0.05),and the rim area,rim volume and disc volume were negatively correlated with MD (all P ＜ 0.05).The ROC curve analysis showed that the largest area under the curve of RNFL thickness in the inferior quadrant of the optic disc region was 0.886,and the specificity and sensitivity was 0.775 and 0.924,respectively.Moreover,the area under the curve of the optic cup/optic disc area was the largest,with sensitivity and specificity of 0.741 and 0.815,respectively.Conelusion OCT for measuring optic disc structure and RNFL thickness can be used for early diagnosis of glaucoma,and it has a high sensitivity and specificity.

OBJECTIVETo evaluate the value of human fatty acid binding protein (h-FABP) in predicting myocardial ischemia and injury in the perioperative period of cardiac surgery, we observed the dynamic changes of h-FABP in perioperative period of patients underwent coronary artery bypass grafting and ventricular septal defects repairing surgery, and evaluated the relationship of h-FABP and ischemia modified albumin (IMA), CK-MB, cTnI.

METHODSPatients underwent coronary artery bypass grafting (n=30) and ventricular septal defect repairing (n=30) surgery between February 2008 and December 2008 were included in this study. Venous blood sample was obtained at preoperative, aortic clamping, aortic unclamping of 10 min, 2 h, 6 h, 12 h, 24 h for the measurements of h-FABP, IMA, cTnI and CK-MB.

RESULTSh-FABP and IMA changed in the same way at various examined time points, h-FABP changes also paralleled cTnI and CK-MB changes, h-FABP peaked early during myocardial ischemia and injury and returned to baseline level at 2 h post myocardial ischemia and injury. Linear correlation analysis showed that the peak value of h-FABP was positively correlated with IMA, CK-MB and cTnI in both CABG group (r = 0.948, 0.964 and 0.961, P < 0.05) and in the VSD group (r = 0.986, 0.978 and 0.957).

CONCLUSIONSh-FABP is an early diagnostic parameter reflecting perioperative myocardial ischemia and injury in cardiac surgery. Quantitative h-FABP monitoring could predict the severity of myocardial ischemia and injury early during cardiac surgery.

Aged ; Albumins ; analysis ; Biomarkers ; blood ; Creatine Kinase, MB Form ; blood ; Fatty Acid-Binding Proteins ; blood ; Humans ; Middle Aged ; Myocardial Ischemia ; diagnosis ; surgery ; Myocardium ; metabolism ; Perioperative Period ; Predictive Value of Tests ; Thoracic Surgery ; Troponin I ; blood

OBJECTIVETo study the distribution of aldehyde dehydrogenase-2 (ALDH2) polymorphisms between healthy Zhuang and Han ethnic individuals in Guangxi Autonomous Region and its influence to the behaviors of alcohol consumption.

METHODSPolymerase chain reaction with confronting two-pair primers (PCR-CTPP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques were used to genotype ALDH2, respectively, and alcohol consumption was recorded in a constructed questionnaire.

RESULTSThe frequencies of ALDH2 alleles (ALDH2(1)/ALDH2(2)) among Zhuang and Han ethnics were 0.511, 0.489 and 0.508, 0.492 respectively (chi2 = 0.001, P > 0.05). The ALDH2 genotypes were verified with PCR-RFLP method. The frequencies of ALDH2(1) genotype in alcoholics (> or = 3 times drinking per week) were 35.59% and 15.67% in Zhuang and Han groups respectively (chi2 = 5.800, P = 0.016).

CONCLUSIONThere was no significant different distribution of ALDH2 genotype among healthy Zhuang and Han ethnic people. The genotype of ALDH2 in different ethnicity might influence individual behavior of alcohol consumption.

Adult ; Aldehyde Dehydrogenase ; genetics ; Aldehyde Dehydrogenase, Mitochondrial ; Alleles ; China ; DNA Primers ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length

OBJECTIVETo investigate the clinicopathological features, diagnosis, treatment and prognosis of primary clear cell carcinoma of the liver (PCCCL).

METHODSThe clinicopathological data of 24 cases with pathologically proven PCCCL in the First Affiliated Hospital of Guangxi Medical University from May 1996 to December 2003 were collected and analyzed.

RESULTSThere were 21 males and 3 females in this group, with an average age of 46 years (range: 30 approximately 78 years). HBV infection was detected in 83.3%, and AFP expression was found in 75.0% of them. Of the 24 cases, 28 tumors were found with an average size of (6.64 +/- 5.54) cm. Liver cirrhosis was found in 75.0% of the patients. Macroscopic and microscopic tumor thrombi were found in 20.8% and 29.2%, respectively. Lymph node metastasis was found in 4.2% of the patents. The 1-, 3-, and 5-year overall survival rates of the 24 cases were 75.0%, 41.7% and 27.8%, respectively, with a median survival time of 29 months.

CONCLUSIONThe clinical characteristics of primary clear cell carcinoma of the liver are similar to that of common hepatocellular carcinoma. It is difficult to be diagnosed preoperatively and final diagnosis depends on pathological examination. Surgical resection is an effective way to achieve favorable treatment outcome and even long-term survival.

Adenocarcinoma, Clear Cell ; blood ; pathology ; surgery ; virology ; Adult ; Aged ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Hepatitis B ; Humans ; Liver Neoplasms ; blood ; pathology ; surgery ; virology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Survival Rate ; alpha-Fetoproteins ; analysis

BACKGROUND AND OBJECTIVESince the proposal of the tumor stem cell hypothesis, considerable interest has been devoted to the isolation and purification of tumor stem cells. Tumor stem cell enrichment from primary tumor derived cell spheres has been demonstrated in specific, serum-free media. This goal of this study is to establish a method of cultivating floating tumor spheres from neuroblastoma cells and to confirm that neuroblastoma spheres are rich in tumor stem cells.

METHODSBone marrow aspirates were obtained from pediatric patients diagnosed with stage IV neuroblastoma. Primary tumor cells were isolated and cultivated in serum-free, stem cell-selective medium. Single sphere-forming cells were cultivated under serum-free conditions; their cloning efficiency and monoclonal tumor sphere formation rates were calculated. The expression of stem cell marker genes Oct-4 and Bmi-1 was detected by RT-PCR in sphere-forming cells and parental neurolastoma cells. Sphere-forming cells were injected into the armpit of nude mice with subsequent assessment for tumor growth. Sphere-forming cells were cultivated in differentiation medium containing 5 μmol/L 13-cis retinoic acid; changes in cell morphology were observed.

RESULTSNeuroblastoma cells formed non-adherent neurospheres under serum-free, stem cell-selective conditions after a period of 4 to 6 days. A single cell dissociated from a neurosphere could reform a monoclonal sphere; cloning efficiency and monoclonal sphere formation rates were 55.3% and 26.3%, respectively. RT-PCR results revealed heightened tumor sphere expression of Oct-4 and Bmi-1 as compared with parental tumor cells. Fourteen days after injection of 10(4) sphere-forming cells into nude mice, a neuroblastoma xenograft formed. Treatment of sphere-forming cells with 13-cis retinoic acid induced a gradual differentiation to neuronal cell morphology.

CONCLUSIONSNeuroblastoma derived tumor spheres enrich tumor stem cells and the cultivation of primary neuroblastoma cells in serum-free, stem cell-selective medium is an effective method to dissociate and purify tumor stem cells in vitro.

Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; drug effects ; Child ; Culture Media, Serum-Free ; Humans ; Isotretinoin ; pharmacology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; metabolism ; pathology ; Neuroblastoma ; metabolism ; pathology ; Nuclear Proteins ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; metabolism ; Repressor Proteins ; metabolism ; Spheroids, Cellular ; pathology ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate effects of intermittent negative pressure on osteogenesis in human bone marrow-derived stroma cells (BMSCs) cultured in vitro.

METHODSThe third passage cells were divided into negative pressure treatment group and control group. The cells in the treatment group were induced by negative pressure intermittently (pressure: 17 kPa, 30 min per time, and four times of each day). The cells in the control group were cultured in conventional condition. The osteogenesis of BMSCs was examined by phase-contrast microscopy. The alkaline phosphatase (ALP) activities were determined. The expression of collagen type I was detected by immunohistochemistry method. The mRNA expressions of osteoprotegerin (OPG) and osteoprotegerin ligand (OPGL) in BMSCs were analyzed by real-time polymerase chain reaction (PCR).

RESULTSBMSCs showed a typical appearance of osteoblast after 2 weeks of induction by intermittent negative pressure. The activity of ALP increased significantly, and the expression of collagen type I was positive. In the treatment group, the mRNA expression of OPG increased significantly (P < 0.05) and the mRNA expression of OPGL decreased significantly (P < 0.05) after 2 weeks, compared with the control. However, 3 days after the exposure to 2-week negative pressure, these were no significantly different from that of the control group (P > 0.05).

CONCLUSIONIntermittent negative pressure could promote osteogenesis in BMSCs in vitro.

Bone Marrow Cells ; physiology ; Cell Culture Techniques ; Collagen Type I ; analysis ; Humans ; Osteogenesis ; Osteoprotegerin ; genetics ; Pressure ; RANK Ligand ; genetics ; RNA, Messenger ; analysis ; Stromal Cells ; physiology

Objective To analyze the results of malaria focus investigation and disposal in Tengchong City,Yunnan Prov-ince between 2015 and 2016,so as to provide evidences for interrupting potential malaria transmission. Methods The malaria foci were investigated and disposed according to the malaria cases reported from"China Information System for Disease Control and Prevention"in Tengchong City between 2015 and 2016. The mosquitoes were captured by mosquito-lured lamp overnight to investigate the malaria vector in local. The peripheral blood samples were collected from the local residents with a febrile history in the foci within two weeks and the accompanists of the cases,and then the rapid malaria diagnosis tests were used to screen the malaria carriers. Results Totally 145 imported malaria cases were reported in Tengchong City between 2015 and 2016,and the focus investigation and disposal rate within 7 d was 100%(145/145). A total of 16186 mosquitoes of 12 species of Anopheles were captured,among which An. sinensis was the predominant,accounting for 64.31%(10410/16186),followed by An. kun-mingensis and An. minimus,with the constituent ratios of 14.15%(2291/16186)and 11.66%(1887/16186),respectively. One person(1.96%,1/51)in the 51 accompanists of the cases was positive in the malaria rapid diagnosis test. Conclusions The malaria transmission vectors such as An. sinensis,An. kunmingensis,An. minimus etc. are still distributed in Tengchong City,and An. sinensis is the predominant species. Therefore,the relevant authorities should continue to strengthen the focus in-vestigation and disposal to interrupt the potential introduced malaria transmission by imported cases in the future.