OBJECTIVEThe expression of C/EBPalpha protein and mRNA during automatically activation process in primary cultures of HSCs were observed in order to explore its possible association with the proliferation and activation of HSCs.

METHODSImmunocytochemistry, Western blot and RT-PCR were used to evaluated the expression of C/EBPalpha protein and mRNA; as well as the expression of alpha-SMA, Desmin, MMP2, type I procollagen (alpha1). The eukaryotic vector harboring the full length cDNA of C/EBPalpha was transfected into activated HSC, then immunocytochemistry was applied to confirm the transfection and evaluate the effect of transfection on the proliferation of HSC by calculating the PCNA-positive cells. The morphological changes of HSC were observed by use of phase-contrast microscope.

RESULTSConstitutive expression of mRNA and protein of C/EBPalpha were detected in primarily cultured HSCs, and the protein was seen in both nuclei and cytoplasm with the latter being dominant. Their expression levels reached highest at day 2 of the culture, then decreased gradually when continually cultured to the day 4, 7, 10, on the other hand, the expression of alpha-SMA, MMP2 and ColI(alpha1) increased steadily. Transient transfection was verified by the fact that much more and stronger C/EBPalpha stain was observed in transfected HSCs than in void-vector transfected cells. In C/EBPalpha gene transfected HSCs, the number of PCNA-positive cells dramatically decreased compared with the void-vector transfected cells 24h after transfection. In addition, the C/EBPalpha gene transfected HSCs died 36 h after transfection, a few surviving cells became longer and thinner in morphology, however the void-vector transfected cells almost all remained alive.

CONCLUSIONSC/EBPalpha was likely involved in the HSCs activation, and over-expressed C/EBPalpha by transfection had inhibitory influence on the proliferation of cultured rat HSCs.

Animals ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; Cells, Cultured ; Collagen Type I ; genetics ; Liver ; cytology ; metabolism ; Male ; Matrix Metalloproteinase 2 ; genetics ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transfection

Human placental decidua basalis-mesenchymal stem cells (PDB-MSCs) are multipotent cells from the human term placenta, which are ethically conducive, easily accessible and high-yielding source. PDB-MSCs can differentiate into adipogenic, osteogenic and neurogenic cells under appropriate conditions, which may be an attractive and alternative source of seed cells for tissue engineering. To investigate the effect of hypoxia (1% O2) on human PDB-MSCs and the expression of cytokine, PDB-MSCs were isolated from human placenta by density gradient centrifugation and cultured in the Dulbecco's modified Eagle's medium-high glucose (DMEM-HG) containing 10% fetal bovine serum (FBS), and the fifth passage of PDB-MSCs were taken. PDB-MSCs were divided into 4 groups according to the concentrations of O2 and FBS: 20% O2, 10% FBS; 20% O2, 0% FBS; 1% O2, 10% FBS; 1% O2, 0% FBS. The proliferation and apoptosis of PDB-MSCs were detected by MTT and flow cytometric analysis at the time points of 6 h, 12 h, 24 h, 48 h, 72 h, and 96 h, respectively. Vascular endothelial growth factor (VEGF) released from PDB-MSCs was detected by enzyme-linked immunosorbent assay (ELISA) at the same time points. The results showed that hypoxia enhanced the proliferation of PDB-MSCs at 12 h under the condition of 10% FBS, while at 24 h under the condition of 0% FBS (P<0.01, n=3). In normoxia, the cells cultured in 10% FBS displayed a significant proliferation compared to those cultured in 0% FBS. However, in hypoxia, the number of cells cultured in 0% FBS (serum deprivation) increased significantly compared to that cultured in 10% FBS at 24 h and 96 h respectively (P<0.05, P<0.01, n=3). With the flow cytometric analysis of cell apoptosis under the condition of hypoxia and serum deprivation, we found that hypoxia and serum deprivation did not induce PDB-MSCs apoptosis (P>0.05, n=3). This conclusion may relate to the expression of VEGF which needs further research. In conclusion, the results obtained indicate that PDB-MSCs are able to bear hypoxia and serum deprivation, suggesting that PDB-MSCs can be used as seed cells for ischemia related tissue engineering.
Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Decidua ; cytology ; Female ; Humans ; Mesenchymal Stromal Cells ; cytology ; Placenta ; cytology ; Pregnancy ; Tissue Engineering ; Vascular Endothelial Growth Factor A ; metabolism

Objective To explore the application of parents' presence induction program (PPIP) during anesthesia induction in children in the operation room and the nursing. Methods 86 children patients (1~11 years old) who would accept the operation were randomly divided into the observation and the control group (n=43 in each group). The observation group was accompanied by their parents when the children accepted the anesthesia induction under the instruction of nurses. But the patients were not present in the control group, and the patients were leaded into the operation room directly by the nurses to accept the anesthesia induction. Blood pressure and pulse changes before and after the anesthesia induction of these children patients were observed. The feeling of their heads of family before the operation and the satisfaction about PPIP were investigated. Results Blood pressure and pulses in the control group were significantly higher than those in the observation group before they were leaded into the operation room (P<0.01). 95% of heads of family hoped to accompany their children when they accepted the anesthesia induction, and 93.7% of heads of family wanted to take part in the PPIP. The satisfaction rate about PPIP was 95% after the completion of PPIP. Conclusions There are many profits about PPIP, such as mitigating the fear and anxiety of children and relaxing the unstable feeling of their heads of families before the anesthesia, and reducing the quantity of pre-anesthetic medications. These are helpful to the anesthesia induction and the cooperation of doctor patients. It is a best anesthesia induction mode worthy to be extended in the operation room.

BACKGROUNDInflammation is a major cause of restenosis after coronary stenting. Intercellular adhesion molecule-1 (ICAM-1) is an important adhesion molecule that plays a key role in the tight adhesion between leukocytes and vascular endothelium. The object of this study was to investigate the association between the K469E polymorphism of the ICAM-1 gene and restenosis after coronary stenting in North Chinese population.

METHODSThe ICAM-1 K469E polymorphism was genotyped using polymerase chain reaction-restriction fragment length polymorphism method in 124 patients who had undergone coronary stenting and coronary angiography at least 3 months earlier. Information on clinical risk factors and procedure-related data were also collected.

RESULTSOf 124 enrolled patients in total, there were 72 cases of in-stent restenosis. The restenosis rate in this population was 58.1%. The frequencies of the three possible genotypes of the ICAM-1 K469E polymorphism were: KK genotype 50.8%, EE genotype 41.9%, and EK genotype 41.9%. Among restenosis patients, the frequency of the KK genotype was 58.3% and the frequency of E allele carriers was 41.7%. Among non-restenosis patients, the frequency of the KK genotype was 40.4%, and the frequency of E allele carriers was 59.6%. The distribution of these two genotype groups between restenosis and non-restenosis patients was significantly different (P = 0.049). Using multivariate logistic regression, the difference between the two groups was more apparent. The odds ratio of KK homozygotes vs E allele carriers was 2.6, with 95% confidence interval 1.2 - 5.8 (P = 0.018). After grading of risk factors, we found that the KK genotype was a stronger predictor of in-stent restenosis in obesity or hyperlipemia patients, with an odds ratio of 9.3 and 3.7, respectively (P < 0.05).

CONCLUSIONIn our study population, KK homozygotes of the ICAM-1 codon 469 mutation had a higher risk of restenosis after coronary stenting, especially in the case of obese or hyperlipemia patients.

Asian Continental Ancestry Group ; genetics ; China ; Codon ; Coronary Restenosis ; genetics ; Female ; Genotype ; Humans ; Hyperlipidemias ; complications ; Intercellular Adhesion Molecule-1 ; genetics ; Male ; Middle Aged ; Obesity ; complications ; Polymorphism, Genetic ; Stents

Adult ; Anesthesia ; methods ; Anti-N-Methyl-D-Aspartate Receptor Encephalitis ; drug therapy ; Cystectomy ; Female ; Humans ; Immunotherapy ; methods ; Young Adult

Objective To test the effects of human-computer interaction intelligent compression package for heparin injection in patients with orthopedic surgery.Methods From July 2015 to June 2016,108 patients with orthopedic surgery were enrolled in the study,and were divided into the experimental group (54 cases) and the control group(54 cases) by random number table.The experimental group was given heparin injection with human-computer interaction intelligent compression package for 3 min,while the control group was given manual compression for 3 min.We evaluated the incidence and severity of subcutaneous hemorrhage and nurses' operating time of two groups.Results The incidence of subcutaneous hemorrhage was 3.9％ in the experimental group,12.4％ in the control group(P＜0.05).The operating time was(100.4±8.7 s) for the experimental group and(233.8±15.3 s) for the control group (P＜0.01).Conclusion Using human-computer interaction intelligent compression package can reduce the incidence of subcutaneous hemorrhage.It can also reduce the working hours of nurses and optimize human resources.

This study was purposed to investigate the relationship between HLA-A, B allele polymorphisms and red blood cell parameters of patients with --(SEA/αα) subtype of α(0)-thalassemia in Han ethnic population of Wuzhou city. The HLA genetic polymorphisms were determined by polymerase chain reaction-sequence-based typing (PCR-SBT) in 57 patients with --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city, Guangxi province, China. Mean corpuscular volume (MCV), hemoglobin (Hb), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were detected by automatic blood analyzer system. HbA2 were detected by electrophoretic method. The statistical analysis was performed by ordinal polytomous logistic regression. The results showed that Hb and HbA2 levels were significantly lower in patients positive for HLA-A*33:03, B*15:01 or B*55:02, and were significantly higher in patients positive for B*15:02 (P < 0.05). It is concluded that several HLA alleles may be associated with Hb level of --(SEA/αα) subtype of α(0)-thalassemia of Han ethnic population in Wuzhou city. This result has the value for understanding phenotype-genotype relationships in thalassemia.
Adolescent ; Adult ; Alleles ; Child ; Child, Preschool ; China ; epidemiology ; Erythrocytes ; cytology ; Ethnic Groups ; genetics ; Female ; Genotype ; HLA-A Antigens ; genetics ; HLA-B Antigens ; genetics ; Hemoglobins, Abnormal ; genetics ; Humans ; Male ; Middle Aged ; Young Adult ; alpha-Thalassemia ; blood ; classification ; epidemiology ; genetics

Objective To observe the association between preprocedural high sensitivity C-reactive protein (hs-CRP) level and incidence of contrast induced acute kidney injury (CI-AKI) in acute coronary syndrome (ACS) patients undergoing percutaneous coronary intervention (PCI) and the impact of atorvastatin pretreatment on CI-AKI. Methods According to the level of preprocedural hs-CRP, 270 ACS patients were divided into three groups: high hs-CRP group (hs-CRP≥3 mg/L, n =176), moderate hsCRP group (hs-CRP 1 -3 mg/L, n =60) and normal hs-CRP group ( hs-CRP ＜ 1 mg/L, n =34).According to the dosage of preprocedural atorvastatin, the high hs-CRP group was further divided into 10 mg group (n =49), 20 mg group (n =66) and 40 mg group (n =61 ). Serum creatinine (Ser), blood urea nitrogen (BUN), cystatin C (Cys C), hs-CRP were measured at before and 24 hours, 48 hours after PCI.CCr and GFR were calculated according to Scr and Cys C. Risk factors for CI-AKI were determined by multivariate logistic regression analysis. Results ( 1 ) Cys C was significantly increased and GFR after PCI significantly reduced in high and moderate hs-CRP groups compared with normal hs-CRP group ( P ＜ 0. 05 ).(2) Incidence of CI-AKI was 43. 18％, 38. 33％ , 20. 59％ in high, moderate and normal hs-CRP groups,respectively (P ＜ 0. 05 ). (3) In high hs-CRP group, postprocedural GFR was significantly higher while postprocedural Cys C and hs-CRP were significantly lower in 40 mg statin subgroup than 10 mg and 20 mg statin subgroups (P ＜ 0. 05), similar trends were documented when comparing 20 mg statin subgroup with 10 mg statin subgroup ( P ＜ 0. 05 ). (4) Multivariate logistic regression analysis showed that pretreatment with high dose atorvastatin was a protective factor for post CI-AKI (20 mg atorvastatin: OR =0. 15, 95％ CI 0. 06 -0. 33, P =0. 00 1 ; 40 mg atorvastatin: OR =0. 10, 95 ％ CI 0. 04 -0. 23, P =0. 001 ), while high levels of preprocedural hs-CRP (OR =2. 06, 95％ CI 1.01 -4. 23, P =0. 048), diabetes mellitus (OR =10.71,95％ CI5.29 -21.70, P=0.001), advanced age (OR =2. 64, 95％ CI 1.05 -6. 63, P =0.038)and renal failure ( OR =5. 14, 95％ CI 1.13 - 23.39, P =0. 034 ) were independent risk factors of CIAKI. Conclusion High hs-CRP level is linked with the development of CI-AKI in ACS patients undergoing PCI and pretreatment with 40 mg atorvastatin is associated with lower incidence CI-AKI, possibly by reducing the postprocedural inflammation responses.

OBJECTIVE: This study aimed to investigate the effect of calcium ion (Ca²⁺) on the migration and osteogenic differentiation of human osteoblasts and explore the proper concentration and correlation mechanism. METHODS: A series of Ca²⁺ solutions with different concentrations was prepared. Osteoblast migration was assessed by Transwell assay, and proliferation was studied via the CCK-8 colorimetric assay. The mRNA expression of osteogenic genes was examined via reverse transcription-polymerase chain reaction (RT-PCR), and the mineralized nodule was examined by alizarin red-S method. After calcium sensitive receptor (CaSR) antagonism, Ca²⁺-induced migration and osteogenic differentiation were analyzed. RESULTS: In the migration experiment, 2, 4, and 6 mmol·L⁻¹ Ca²⁺ could promoted osteoblast migration at three timepoints (8, 16, and 24 h), whereas 10 mmol·L⁻¹ Ca²⁺ considerably inhibited migration at 8 h. The Ca²⁺ concentration range of 2-10 mmol·L⁻¹ could promote proliferation, osteogenic differentiation, and mineralization of human osteoblasts. Moreover, mineralization was predominantly induced by 8 and 10 mmol·L⁻¹ Ca²⁺. CaSR antagonism could reduce Ca²⁺-induced migration and osteogenic differentiation of human osteoblasts. CONCLUSIONS Low Ca²⁺ concentration favored osteoblast migration, whereas high Ca²⁺ concentration favored osteogenic differentiation. The Ca²⁺ concentrations of 4 and 6 mmol·L⁻¹ could substantially induce osteoblast migration and osteogenic differentiation, and the Ca²⁺-CaSR pathway participated in signal transduction.
Calcium ; physiology ; Cell Differentiation ; Cell Movement ; Cell Proliferation ; Humans ; Osteoblasts ; Osteogenesis ; physiology ; Signal Transduction

Introduction:Patients with metastatic nasopharyngeal carcinoma (NPC) have variable survival outcomes. We have previously shown that an elevated peripheral blood lymphocyte-to-monocyte ratio (LMR) is associated with an increased metastatic risk in patients with primary NPC. The present study aimed to investigate the prognostic value of pretreatment LMR in a large cohort of metastatic NPC patients. Methods:Clinical data of 672 patients with metastatic NPC diagnosed between January 2003 and December 2009 were analyzed. The peripheral lymphocyte and monocyte counts were retrieved, and LMR was calculated. Receiver operating characteristic (ROC) curve analysis and univariate and multivariate COX proportional hazards analyses were performed to evaluate the association of LMR with overall survival (OS). Results:Univariate analysis revealed that an elevated absolute lymphocyte count (≥1.390 × 109/L) and LMR (≥2.475) as well as a decreased monocyte count (<0.665 × 109/L) were significantly associated with prolonged OS. Multivariate Cox proportional hazard analysis showed that LMR (hazard ratio [HR]=0.50, 95%confidence interval [CI]=0.41–0.60, P<0.001), absolute lymphocyte count (HR=0.77, 95%CI=0.64–0.93, P=0.007), and monocyte count (HR=1.98, 95%CI=1.63–2.41, P<0.001) were independent prognostic factors. By stratification analyses, only LMR remained a significant predictor of prognosis. Conclusion:We identified pretreatment LMR as an independent prognostic factor for patients with metastatic NPC. Independent validation of our findings is needed.