BACKGROUND:We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cels at-80℃.
OBJECTIVE:To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cels and to verify the biological features of bone marrow mesenchymal stem cels after long-term cryopreservation.
METHODS: Bone marrow mesenchymal stem cels were cultured using adherent method and the biological features and purity of cels were detected using immunofluorescence method. Bone marrow mesenchymal stem cels were cryopreserved in the cryoprotectant medium containing low-sugar DMEM, fetal bovine serum and dimethyl sulfoxide at different proportions at-80℃ for a short term. Then, the optimal cryoprotectant was selected to storage the bone marrow mesenchymal stem cels. After 1, 3, 6 months of cryopreservation, the cels were resuscitated, cultured and passaged. Passage cels were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cels cryopreserved at-80℃.
RESULTS AND CONCLUSION:Cryoprotectant medium of 80% DMEM+10% fetal bovine serum+10% dimethyl sulfoxide was suitable for cryopreserving MSCs at -80℃, and resuscitated cels were able to proliferate in vitro, and passage normaly, indicating the cryopreserved bone marrow mesenchymal stem cels stil maintain the original biological activity.

Adult ; Alkaline Phosphatase ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervix Uteri ; metabolism ; pathology ; surgery ; Choriocarcinoma ; metabolism ; pathology ; Chorionic Gonadotropin ; metabolism ; Diagnosis, Differential ; Female ; GPI-Linked Proteins ; metabolism ; Humans ; Hysterectomy ; Placental Lactogen ; metabolism ; Pregnancy ; Trophoblastic Tumor, Placental Site ; metabolism ; pathology ; surgery ; Uterine Cervical Neoplasms ; metabolism ; pathology ; surgery

Brain magnetic resonance imaging (MRI) of the elderly often reveals white matter changes (WMCs) with substantial variability across individuals.Our study was designed to explore MRI features and site-specific factors of ischemic WMCs.Clinical data of consecutive patients diagnosed with ischemic cerebral vascular disease who had undergone brain MRI were collected and analyzed.Multi-logistic regression analysis comparing patients with mild versus severe WMCs was performed to detect independent associations.Analyses of variance (ANOVAs) were used to detect regionally specific differences in lesions.We found that lesion distribution differed significantly across five cerebral areas,with lesions being predominant in the frontal lobe and parieto-occipital area.To explore WMCs risk factors,after adjusting for gender,diabetes mellitus,and hypertension,only age (P＜0.01),creatinine (P=0.01),alkaline phosphatase (ALP) (P=0.01) and low-density lipoprotein cholesterol (LDL-C) (P=0.03) were found to be independently associated with severe WMCs.Age (P＜0.001) was strongly associated with WMCs in the frontal lobe while hypertension was independently related to lesions in the basal ganglia (P=0.048) or infratentorial area (P=0.016).In conclusion,MRI of WMCs showed that ischemic WMCs occurred mostly in the frontal lobe and parieto-occipital area.The infratentorial area was least affected by WMCs.Typically,age-related WMCs were observed in the frontal lobes,while hypertension-related WMCs tended to occur in the basal ganglia and infratentorial area.

Objective:To investigate the effect of TPF-DM on blood glucose levels and immune function in severe traumatic brain injury (STBI) patients.Methods:We consecutively included 60 STBI patients who were randomly divided into control and experimental groups.The two groups of patients were treated with TPF and TPF-DM as the main energy supply,respectively.Total protein,albumin,prealbumin,procalcitonin (PCT),immunoglobulinM (IgM),immunoglobulinG (IgG),immunoglobulinA (IgA) and blood glucose of all patients were detected on day 1,7 and 14.Immune function related indicators,serum protein and blood glucose were statistically analyzed.Results In the first week,the mean blood glucose level and incidence of hypoglycemia were not significantly different between the two groups (P ＞ 0.05).However,the blood glucose variability (BGV) was significantly different (P ＜ 0.05).In the second week,both the blood glucose level and BGV were in normal range with no significant difference between two groups (P ＞ 0.05).At dayl,the total protein,albumin,prealbumin,IgM,IgG and IgA of all the patients were below the normal level,while the PCT was exact opposite,but with no significant difference between the two groups (P ＞ 0.05).At day7,PCT decreased compared to dayl in both the control and experimental groups,while the total protein,albumin,prealbumin,IgM,IgG and IgA increased,but the experimental group increased much more compare to the control group (P ＜ 0.05).At dayl4,all these indexes returned to normal levels with no significant difference between the two groups (P ＞ 0.05).Conclusion:TPF-DM is significantly better than TPF to control the blood glucose level.TPF-DM may have a positive effect on the control of early excessive inflammation and infection,and then improve the immune function.Yet the potential physiopathologic mechanism needs further study.

Objective To investigate the value of perfusion imaging in predicting the status of isocitrate dehydrogenase 1 (IDH1) in glioblastoma.Methods Retrospectively studied 30 patients with glioblastoma multiforme with wild type IDH1 (IDHw) and 30 patients with mutant IDH1 (IDH1m) in Department of Neurosurgery,Shanxi Provincial People's Hospital from September 2014 to February 2016.Ktrans and Ve within the enhancing portion of each tumor were measured by using DCE-MRI data.rCBF and rCBV within the enhancing portion of each tumor were measured by using DSC-MRI.Four parameters were represented as (x) ± s,each of the 4 parameters was compared between patients with wild type IDH1 and mutant IDH1 by using the t-test.The performance in discriminating between the two entities was evaluated by using receiver operating characteristic analysis.Results Ktrans,Ve,rCBF and rCBV inside the enhancing lesion were significantly higher in patients with wild type IDH1 than in those with mutant IDH1.There was a statistically significant difference between IDH1w group and IDH1m group of rCBF value (P ＜ 0.05).The area under the curve for Ktrans,Ve,rCBF,and rCBV inside the enhancing lesion were 0.850,0.873,0.739 and 0.772,respectively.The value of rCBF value of the Ktrans value Ve value of IDH1w was significantly higher than that of IDH1m(P ＜0.05) in the value of the Ktrans value Ve value of rCBF,the value of the AUC value in rCBV was not significantly different with the combination of the 4 parameters of the diagnostic performance (AUC =0.915).Conclusions Ktrans,Ve,rCBF and rCBV calculated from MRI are useful for predicting the IDH1 mutation status.This method is not only for the classification of brain glioblastoma diagnosis has important value,and in glioblastoma classification is of important value in the preoperative noninvasive evaluation,and it has reference significance for predicting the prognosis of patients.

Haff disease is a type of human rhabdomyolysis characterized by the sudden onset of unexplained muscular rigidity and an elevated serum creatine kinase level within 24 h after consuming cooked aquatic products.Here,we reviewed a previous study on Haff disease and summarized the clinical manifestations,epidemiological characteristics,and etiological data to confirm the incidence and global epidemiology of the disease and identify the most common seafood vectors.Future directions for Haff disease study will include further prospective etiological studies and the development of prevention and control strategies.

OBJECTIVETo investigate the possible association between ZNF230 gene and azoospermia.

METHODSScreening for mutation of all 6 exons of ZNF230 gene was performed by denaturing high performance liquid chromatography(DHPLC) in 99 patients with azoospermia and in 115 healthy men as controls.

RESULTSAn A-->G transition at nucleotide 316 in exon 6 was identified. There were significant differences in the distribution profiles of both allele and genotype frequencies between patient group and control group (P < 0.01 and P < 0.05, respectively). In addition,there was a statistically significant difference in the serum follicle stimulating hormone (FSH) level between the patients with GG/GA genotype and those with AA genotype (P < 0.05).

CONCLUSIONZNF230 gene may be associated with azoospermia, and the A316G mutation may be correlated with the serum FSH level.

Adult ; Azoospermia ; diagnosis ; genetics ; Base Sequence ; Chromatography, High Pressure Liquid ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Gene Frequency ; Genetic Testing ; Genotype ; Humans ; Male ; Mutation ; Polymerase Chain Reaction ; Transcription Factors ; genetics ; Young Adult

OBJECTIVEMajor histocompatibility complex class I C-related molecules A and B (MICA and MICB) are innate immune system ligands for the NKG2D receptor expressed by natural killer cells and activated CD8(+)T cells. Our previous study showed that 5-aza-2'-deoxycytidine (5-aza-dC), a DNA methyltransferase inhibitor, can induce the expression of MICB and sensitized cells to NKL-cell-mediated cytolysis. The aim of this study was to determine the expression level of MICA in HepG2 cells (an HCC cell line) and L02 cells ( a normal liver cell), and to investigate the effect of 5-aza-dC on MICA expression in HepG2 cells.

METHODSCells were treated with 5-aza-dC, caffeine and ATM-specific siRNA. The cell surface MICA protein on HepG2 cells and L02 cells was determined using flow cytometry. The mRNA level was detected using real time RT-PCR.

RESULTSMICA was undetectable on the surface of L02 cells, but was highly expressed on HepG2 cells. MICA expression was upregulated in response to 5-aza-dC treatment (P less than 0.05), and the upregulation of MICA was partially prevented by pharmacological or genetic inhibition of ataxia telangiectasia mutated (ATM) kinase (P less than 0.05).

CONCLUSIONOur data suggest that 5-aza-dC induces the expression of MICA by a DNA damage-dependent mechanism.

Ataxia Telangiectasia Mutated Proteins ; Azacitidine ; analogs & derivatives ; pharmacology ; Caffeine ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; Cell Cycle Proteins ; antagonists & inhibitors ; metabolism ; Cell Line ; Cell Membrane ; metabolism ; DNA Damage ; DNA-Binding Proteins ; antagonists & inhibitors ; metabolism ; Flow Cytometry ; Hep G2 Cells ; Hepatocytes ; metabolism ; Histocompatibility Antigens Class I ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; Protein-Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Proteins ; antagonists & inhibitors ; metabolism ; Up-Regulation

BACKGROUNDEquatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber cells. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens.

METHODSLens epithelia were dissected into central (

RESULTSBy microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three (mitogen-activated protein kinase 1 (MAPK1), nidogen (NID), small nuclear ribonucleoprotein N (SNRPN)) out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor beta (TGFbeta) signaling and glycolysis pathways.

CONCLUSIONThe results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells.

Animals ; Cattle ; physiology ; Electrophoresis, Gel, Two-Dimensional ; Epithelium ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; In Vitro Techniques ; Lens, Crystalline ; metabolism ; Oligonucleotide Array Sequence Analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

To evaluate the inhibitory effects of long antisense RNA on HBV replication in HepG2.2.15 cells. The coding region of HBV S gene was cloned into pTARGET vector in sense and antisense orientations and the recombinant plasmids were transfected into HepG2.2.15 cells which were divided into HBS2 (antisense RNA) group, HBS4 (sense RNA) group and control group. HBsAg and HBeAg in the culture supernate were detected by ELISA. The HBV DNA in the supernate was quantified by real-time PCR. After treatment, the levels of HBsAg in HepG2.2.15 cell supernatants of three groups were 0.621+/-0.027, 3.399+/-0.018 and 2.232+/-0.187 respectively; the levels of HBeAg were 0.749+/-0.019, 1.548+/-0.025 and 1.570+/-0.044 respectively and the levels of HBV DNA were 1.597+/-0.082, 3.381+/-0.297 and 3.610+/-0.063 respectively. The expressions of HBsAg and HBeAg and the HBV DNA level in HBS2 group were remarkably reduced as compared to the control (Z = -2.309, P value is less than 0.05); whereas the sense plasmid transfection (HBS4) did not affect HBeAg (Z = -0.866) and HBV DNA (Z = -1.155) levels in the culture supernate but slightly increased the HBsAg level (Z = -2.309). Antisense RNA might be a useful tool to repress HBV replication.
DNA, Viral ; genetics ; Gene Expression Regulation, Viral ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B Surface Antigens ; analysis ; Hepatitis B e Antigens ; analysis ; Hepatitis B virus ; genetics ; physiology ; Humans ; Plasmids ; RNA Interference ; Transfection ; Virus Replication ; genetics