BACKGROUND:Fibroblast growth factor can promote proliferation of mesenchymal stem cells (MSCs).grew along the wall.However,there are few reports on the differentiation of MSCs into hepatocytes following fibroblast growth factor induction.When mass concentration of hepatocyte growth factor was 1 μg/L,it can promote mitosis of hepatocytes,and is the strongest mitogenic agent for normal hepatocytes.OBJECTIVE:To explore the biological characteristics of human umbilical cord MSCs/n vitro and their differentiation ability to hepatocyte-like cells under the induction of chemical factors.DESIGN,TIME AND SETTING:The cytological in vitro study was conducted at the Institute of Blood,Jinan University from August 2008 to April 2009.MATERIALS:Umbilical cord was obtained from healthy full-term fetus,which was provided by the Guangzhou Huaqiao Hospital.The parturient signed informed consent.Hepatocyte growth factor and flbroblast growth factor were bought from Peprotech,USA.METHODS:The MSCs from human cord were isolated and cultured by type Ⅳ colagenase digestion+differential adherence.At the third passage,MSCs received cell surface antigen analysis and cell cycle determination to detect their ability to differentiate into adipocytes and osteoblasts.At the fifth passage,MSCs were adjusted into 5×10~9/L and assigned into 2 groups.BMSCs in the control group were incubated in DMEM/F12 containing 5% fetal bovine serum.BMSCs in the induction group were treated with above-mentioned medium supplemented with 20 μg/L hepatocyte growth factor and 10 μg/L flbroblast growth factor.MAIN OUTCOME MEASURES:The following parameters were measured:biological characteristics of human umbilical cord MSCs and differentiation of human umbilical cord MSCs into hepatocyte-like cells in vitro.RESULTS:At the third passage,MSCs derived from human cord expressed CD29,CD44,CD105,but not antigens of hematopoietic CD34,CD45,and 92.2% of them were in G_0/G_1 phase.Oil red O staining showed cytoplasm presented red granules.Alkaline phosphatase staining demonstrated that cytoplasm was black,with the differentiation ability into adipocytes and osteoblasts.Following 10 days of combined induction of hepstocyte growth factor and fibroblast growth factor,RT-PCR and Western blot results confirmed that cells expressed alpha fetoprotein and albumin.Negative expression was found in the control group.CONCLUSION:Human umbilical cord contained plenty of MSCs,with strong potential of multi-differentiation.Umbilical cord MSCs can differentiate into hepatocyte-like cells following combined induction of hepatocyte growth factor and fibroblast growth factor.

BACKGROUND: The relationship between cellular morphosis and function is undetachable, but there is little investigation about ultrastructure of human umbilical cord mesenchymal stem cells (hUCMSCs).OBJECTIVE: To study the relationship between the functions of hUCMSCs and their ultrastructure obtained by atomic force microscope (AFM). METHODS: hUCMSCs were isolated, cultured, expanded after enzyme digestion. P3 cells were observed under AFM. Immunophenotype and cell cycle were analyzed by flow cytometry, as well as induction of the adipogenic, osteogenic differentiation of hUCMSCs were identified using Oil red O staining and alkaline phosphatase staining. RESULTS AND CONCLUSION: hUCMSCs at passage 3 were strongly positive for CD44 and CD29, weakly positive for CD106, but negative for hematopoietic marker CD34. Cells in G_0/G_1 phase accounted for 80%. Proliferation index was 19.9%. Following adipogenic induction, alkaline phosphatase staining demonstrated brown cytoplasm in cube and polygonal cells. AFM showed hUCMSCs were spindle shape, obvious cytoskeletal filament that connected into nets, which fit for the strong capacities for proliferation, migration and differentiation.

Objective:To investigate the quantification of CD4~+CD25~+ regulatory T cells and distribution of CD4~+CD8~+ T lymphocyte subgroup in peripheral blood of patients in chronic hepatitis B (CHB),and to reveal relationship between CD4~+CD25~+ regulatory T cells,CD4~+CD8~+ T lymphocyte subgroup and HBV infetion as well.Methods:CD4~+CD25~(high),CD4~+CD25~+Foxp3~+Treg and CD3~+CD4~+CD8~+T lymphocyte subgroup in peripheral blood from 50 patients with CHB and 20 healthy controls was analyzed using flow cytometry.HBV DNA was detected by fluorescence quantitative PCR.Results:The number of CD4~+CD25~(high)Tregs in patients with CHB was obviously higher than that in healthy controls(P＜0.01)and increased with copies of HBV DNA.The same with the change of CD4~+CD25~+Foxp3~+Tregs in patients with CHB and there was a positive correlation between CD4~+CD25~(high)Tregs and CD4~+CD25~+Foxp3~+Tregs(r=0.890,P＜0.001).Compared with healthy controls,the frequency of CD4~+T cells and the ratio of CD4~+/CD8~+ in patients with CHB was declined,but there was no significant difference in the frequency of CD3~+T cells and CD8~+T cells between them(P＞0.05).The variation in the number of CD4~+CD25~(high)Tregs was correlated positively with the copies of HBV DNA(r=0.782,P＜0.001)and glutamic-pyruvic transaminase(ALT)(r=0.432,P＜0.005)separately,but negatively with the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+(P＞0.05).The variation in the frequency of CD3~+,CD4~+,CD8~+T cells and the ratio of CD4~+/CD8~+ was also correlated negatively with the copies of HBV DNA(P＞0.05).Conclusion:The number of CD4~+CD25~(high)Tregs increases in patients with CHB and is in accordance with the copies of HBV DNA and increased level of ALT.Further studies should be done to investigate weather CD4~+CD8~+ T lymphocyte subgroup could be used to monitor the state of community.

ObjectiveObjective To investigate the value of pancreatic elasticity based on shear wave elastography (SWE) in predicting the risk of pancreatic fistula, to measure pancreatic hardness by determining the SWE level of the pancreatic body, and to reduce the development of pancreatic fistula after pancreaticoduodenectomy. MethodsA retrospective analysis was performed for the clinical data of 53 patients who were admitted to The Affiliated Hospital of Binzhou Medical University from October 2017 to February 2019 and underwent pancreaticoduodenectomy (PD). The 53 patients were divided into pancreatic fistula group with 10 patients who developed pancreatic fistula after PD and non-pancreatic fistula group with 43 patients who did not develop this disease after PD. The elasticity value of the pancreatic body measured by SWE was used to reflect the tissue elasticity of the pancreas and evaluate the hardness of the pancreas. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between groups. A logistic regression analysis was used for univariate analysis, and the Spearman’s correlation coefficient was used to investigate the correlation between SWE and other laboratory data. ResultsBody mass index (BMI) (t=1.321), preoperative total bilirubin (t=1.347), diameter of the main pancreatic duct (t=1.385), maximum SWE value (t=1.728), mean SWE value (t=1.634), and intraoperative pancreatic hardness (χ2=4.983) were risk factors for pancreatic fistula (all P＜0.05). Maximum SWE value and mean SWE value were negatively correlated with age and time of operation (maximum SWE value: r=-0.329 and -0.260, both P＜0.05; mean SWE value: r=-0.282, and -0.282, both P＜0.05) and positively correlated with BMI (r=0.275 and 0.350, both P＜0.05). ConclusionSWE level is an independent risk factor for pancreatic fistula after pancreaticoduodenectomy, and the SWE level of the pancreatic body has a high predictive value and guiding significance in surgical treatment.

Objective To explore the effects and the possible molecular mechanism of flavonoids of puerarin (PR) on chronic myelogenous leukemia (CML) cell line K562 and acute promyelocytic leukemia (APL) cell line NB4 in vitro. Methods MTT assays were used to detect the inhibitory effects of cell proliferation. The apoptosis of K562 and NB4 cells was detected by flow cytometry marked with Annexin V/PI. The expression of bcr-abl, p53, bcl-2, Fas/FasL in K562 cells and JNK, PARP, bcl-2 and Caspase 3 in NB4 cells at protein level was detected by Western blot. Results PR could inhibit the proliferation of K562 and NB4 cells in a time-dose dependent manner. The expression of protein levels of bcr-abl fusion gene declined, while the p53 protein otherwise increased, and both were in a dose-dependent manner (F = 18.74, P <0.05). The application of PR had no effect on bcl-2 and Fas/FasL protein expression in K562 cells. The JNK, PARP and Caspase3 proteins were upregulated in NB4 cells, while bcl-2 was downregulated with the increasing concentrations of PR (F=42.32, P <0.05). Conclusion PR could inhibit leukemic cell proliferation, induce cell cycle block, and increase cell apoptosis through different molecular mechanisms. It suggestes that PR might potentially be a kind of broad spectrum anti-leukemia agent.

Objective To compare acoustic radiation force impulse(ARFI)and supersonic shear imaging (SWE) in diagnosis of liver fibrosis in patients with chronic hepatitis B. Methods Eighty patients with chronic hepatitis B having underwent ARFI and SWE examination were enrolled in this study.The elastic modulus E value(EI)was measured by SWE.The liver shear wave velocity(VTQ)was measured by ARFI.All patients underwent liver biopsy.The diagnostic values of SWE and ARFI for liver fibrosis were analyzed with Sperman correlation and the ROC curve.Results The values of EI and VTQ were increased with the pathological stage determined by liver biopsy and there were significantly differences(P<0.01).The correlation coefficient of SWE and ARFI was 0.651,P<0.01.The correlation coefficient of SWE, ARFI and pathological stage determined by liver biopsy were 0.784 and 0.683 and there were significant differences(P<0.01).The areas under ROC for diagnosing liver fibrosis≥S2,≥S3 and =S4 by using SWE were 0.912, 0.934 and 0.955 respectively and those by using ARFI were 0.870, 0.892 and 0.884. The sensitivity of ARFI in diagnosing liver fibrosis was similar with SWE, but SWE showed higher specificity (Z=8.756,P < 0.01; Z=10.802,P < 0.01; Z=15.871,P < 0.01). Conclusions Both SWE and ARFI can be effectively used in the evaluation of liver fibrosis in patients with chronic hepatitis B.The SWE technology has more advantages.

Objective To study the prevalence and related risk factors of type 2 diabetes with coronary heart disease(CHD) and cerebral infarction after long-term treatment.Methods Two hundred and eighty-five patients with type 2 diabetes who had a hospitalization interval for four to twelve years between the first and second hospital stays were included in group A.Among them,85 patients (about 4 to 7 years) with a hospitalization interval about 5 years (4 to 7 years) between the first and second hospital stays(group B).100 patients with a hospitalization interval about 10 years (8 to 12 years) between the first and second hospital stays(group C).Blood pressure,blood glucose,HbA1c,blood lipids,myocardial enzymology,electrocardiogram,coronary angiography,cranial CT or MRI were tecorded.The prevalence of CHD and cerebral infarction were compare,regression analysis was conducted between coronary heart disease,cerebral infarction and the risk factors.Results DBP,FBG,PBG2h,TC,LDL-C of the second hospitalization were significantly lower than those of the first hospitalization.The morbidity of CHD and cerebral infarction increased along with the extension of the course.The morbidity of cerebral infarction in CHD patients were significantly higher than those in non-CHD patients with the interval about 10 years.Logistic regression analysis proved that age was risk factor and HDL-C was protective factor of CHD,SBP,FBG were risk factors of cerebral infarction.Conclusion The morbidity of CHD and cerebral infarction increased along with the extension of the course and there were significantly relationship between CHD and cerebral infarction in T2DM patients after about 10 years treatment.

Objective To explore the monitoring of cerebrospinal fluid (CSF) dynamics in a model of brain herniation induced by acute intracranial hypertension in Guangxi Bama-Mini pigs by phasecontrast cine magnetic resonance imaging (PC cine MRI).Methods Femoral artery blood were extracted from 10 pigs,and injected into the frontal and temporal parietal lobe to make a model of brain herniation induced by acute intracranial hypertension.The mean arterial blood pressure (MAP),intracranial pressure (ICP),and cerebral perfusion pressure (CPP) were monitored.Routine T1WI,T2WI,coronal,sagittal and cerebrospinal fluid flow sequence (fast PC cine slice) which positioned on the cervical 3 (C3) vertebral body as the center and perpendicular to the spinal scans were performed on all experimental animals before and after blood injection with 3.0T Magnetic Resonance Imaging.The ICP,MAP,CPP,the absolute values of CSF peak flow velocity and the absolute value of carotid peak flow velocity before and after blood injection were compared.Results The ICP,MAP,CPP,and the absolute value of CSF peak flow velocity before injection of autologous arterial blood were statistically significant as compared with those after blood injection [(6.80±2.044) mmHg vs (52.20±1.619) mmHg,(76.80±7.068) mmHg vs (142.80±12.399) mmHg,(70.00±6.074) mmHg vs (90.50±12.250) mmHg,and the absolute value of CSF peak flow velocity was (243.20±77.671) mm/s vs (201.40±55.482) mm/s,respectively,P＜0.01].The absolute value of the peak velocity of the carotid artery before blood injection was not statistically significant compared with that after blood injection [(876.80±239.908) mm/s vs (799.40±241.829) mm/s,P＞0.05].Conclusion After the formation of brain herniation induced by acute intracranial hypertension,the CSF flow in the C3 level spinal canal showed a low dynamic change,and the CSF flow velocity waveform was disordered and malformed.The non-invasive measurement of CSF dynamics by PC cine MRI can provide an important basis for the change of CSF dynamics in the model of brain herniation induced by acute intracranial hypertension,and provide a theoretical basis for further research on damage control neurosurgery in the future.

Objective:To evaluate the effects of learning initiative on teaching effectiveness.Methods:The research subjects were the 2nd year medical students of Tongji Medical College of Huazhong University of Science and Technology. The learning initiative factor was calculated from the data of the attendance registration of the on-line learning of Physiology, and the performance of the on-line test was used as an evaluation indicator of teaching effectiveness. SPSS software was used to perform correlation analysis between the learning initiative factor and teaching effectiveness. Results:We found that learning initiative could significantly affect the teaching effectiveness, with differences among different specialties. There was a positive correlation between learning initiative and teaching effectiveness in clinical and pediatric medicine, while no correlation was observed in preventive medicine and medical imaging.Conclusion:In conclusion, learning initiative can affect teaching effectiveness, and the intensity of this effect shows difference among different specialties.

Objective To investigate the clinical characteristics,antibiotic susceptibilities and serotypes of Group B Streptococcus(GBS)isolated from neonatal meningitis to provide references for the prevention and treatment of neonatal GBS meningitis. Methods From June 2013 to June 2016,we surveyed the GBS strains iso-lated from purulent meningitis of < 90 days infants from Guangzhou Women and Children′s Medical Center. The GBS isolates were identified and the minimum inhibitory concentration of the antibiotics was determined by Vitek 2 Compact automatic bacterial identification system.GBS serotyping was performed using Strep-B-Latex?rapid latex agglutination test kit. Results A total of 46 cases of neonatal GBS meningitis,15 cases of early-onset infection and 31 cases of late-onset infection were diagnosed. 78.3% of GBS meningitis with varying degrees of complica-tions.Among 41 survivors with 3~24 months follow-up,50% of the early-onset and 44.8% of the late-onset GBS meningitis with varying degrees of neurological sequelae.Four capsular types were identified among the 46 isolates, serotype-Ⅲwas the most prevalent(73.9%),followed by Ib(19.6%),V(4.3%)and Ia(2.2%).All the isolates were susceptible to penicillins,cephalosporins,linezolid and vancomycin. Conclusion The highly pathogenic serotype-Ⅲ was the predominant serotype among neonatal GBS meningitis in Guangzhou,Therefore,it is neces-sary to strengthen the epidemiological surveillance of GBS invasive infection and the effective implementation of pre-ventive measures.