Objective To investigate whether hepatocyte growth factor ( HGF ) and insulin-like growth factor (IGF1) induce cardiac stem cells (CSCs) to proliferate and directly differentiate into cardiomyocytes in vitro.Methods The myocardial tissues were dissected for primary culture of CSCs with the method of explants .The expressions of c-kit and CD34 were examined with immunofluorescence .Primary cells were purified with c-kit by flow cytometry.CFDA SE fluorescent probe was used to detect the proliferation of c-kit+CSCs.C-kit +CSCs were divided into two groups , and cardiac stem cells group and co-cultured with cardiomyocytes group , both group were cultured with HGF and IGF 1.An inverted microscope was used to observe changes in cell number and morphology in different periods .Living cells workstation was used to observe CFDA SE fluorescence intensity , to acquire images and do statistical analysis .Immunofluorescence technique was used to detect the expression of Nkx 2.5 and cardiac troponin T .Results In cardiac stem cells group ,CSCs had no obvious changes in cell number .In co-cultured with cardiomyocytes group , CSCs proliferated and had changes in morphology .Nkx2.5 and cTnT were positively expressed . Several CSCs differentiated into beating cardiomyocytes . Conclusion In co-cultured with cardiomyocytes condition , HGF and IGF1 may promote CSCs to proliferate and differentiate into beating cardiomyocytes .

This article was aimed to study the protection effects of Pa-Bing Formula No. 2 (PBFN-2) on neurons of substantia nigra in Parkinson's disease (PD) rats models in order to explore the possible mechanism of PBFN-2 in PD treatment. Rats were stereotaxically injected with 6-hydroxydopamine (6-OHDA) solution into the left stria-tum in two-site. Rat showed consistent right whirling and the number of rotation was more than 7 r·min-1 induced by APO in 30 min, then the rat was judged as PD model. A total of 14 rats modeled successfully were randomly di-vided into the model group and the treatment group. At the same time, the normal group and sham operation group were also established. Same volume of distilled water was given to the normal group, sham operation group and the model group. PBFN-2 (32.0 g·kg-1) was given to the treatment group for 4 weeks. Microscope was used to observe pathological changes of substantia nigra by nissl stain and changes of TH, Nrf2 and HO-1 immunohistochemical stain. The results showed that compared with the normal group, the nissl bodies were badly injured. Expressions of TH-positive cells were obviously reduced (P<0.01). The expression of Nrf2 nucleus protein and HO-1 protein were obviously increased in substantia nigra of PD rats in model group (P<0.01). Compared to the model group, PBFN-2 effectively increased nissl bodies in neuronal cells of substantia nigra of PD rats, and elevated the number of TH-immunoreactive cells in substantia nigra (P<0.05). The expressions of Nrf2 nucleus protein, HO-1 protein were significantly up-regulated (P<0.05). It was concluded that PBFN-2 had an obvious neuroprotection on the neuronal cells in substantia nigra of PD rats induced by 6-OHDA. The underlying mechanisms may be associated with regu-lation of Nrf2 nucleus protein and HO-1 protein expressions.

OBJECTIVE:To discuss the drug resistance situation and influential factors of sputum culture positive tuberculosis patients in a specialized subject hospital. METHODS:The information of sputum culture positive tuberculosis inpatients were col-lected from Heze infectious disease hospital during Jan. 1st 2014 to Apr. 30th 2015;the results of drug sensitive test were analyzed retrospectively by using absolute concentration method. RESULTS:Among 259 patients,137 cases were drug-resistant(52.90%), among including 69(26.64%) mono-drug resistant cases,52(20.08%) poly-drug resistant cases,15(5.79%)multi-drug resistant cases and 1(0.39%)extensively drug resistant case;streptomycin (31.66%) and amikacin (15.44%) were drugs with the highest drug-resistant rate;there was no statistical significance in resistant rate between different genders and ages(P>0.05),the rate of drug resistance in first-treatment patients was 47.21%,and that of the re-treatment patients was 70.97%,the difference was statisti-cal significant(P<0.05). In addition to ethambutol and kanamycin,other all had mono-drug resistance,there were 27 combinations of poly-drug resistance and 9 combinations of multi-drug resistance. Logistic regression analysis showed that treatment history was a risk factor for drug-resistance,the risk of re-treatment patient was 2.734 times of first-treatment patients. CONCLUSIONS:Sputum culture positive tuberculosis inpatients patients have high drug resistance and complex spectrum in the hospital;we should pay great attention to the risk of drug resistance in senile patients and re-treatment patients,and rational use of streptomycin,amikacin,isoni-azid and capreomycin.

Aim To construct and express the eukary-otic expression vector of double-stranded RNA-depend-ent protein kinase (PKR)fusion green fluorescent and analyse its antiviral activity of HBV in vitro.Methods The PKR gene was cloned into an empty expression vector pEGFP-N1 using molecular clone technology. After being confirmed by restriction enzyme digestion and sequencing methods,the recombinant plasmid was named as pEGFP-PKR that was subsequently transfect-ed into HepG2.2.15 cells using LipofectamineTM2000. The expression level of PKR in HepG2.2.15 cells was confirmed by using fluorescent microscopy. Mean-while,HBV DNA and HBsAg/HBeAg were detected by real-time PCR and electrochemiluminescence meth-od,respectively.Results Both restriction enyme di-gestion and sequencing assays showed that the recombi-nant vector pEGFP-PKR was successfully constructed in our study.Fluorescent microscopy observation indi-cated that the fusion protein pEGFP-PKR expressed ef-ficiently in HepG2.2.15 cells.Moreover,compared with the empty vector group,the expression of HBV antigen in supernatants was significantly decreased (P<0.05 ).However,the extracellular HBV DNA ex-pression was not inhibited significantly.Conclusion In vitro,PKR proteion has certain antiviral activity of HBV.

The effect of cholesterol ester transfer protein(CETP) on SR-B1 mRNA expression in mouse liver was investigated.The results demonstrated that CETP transgenic mice showed lower serum total cholesterol and high density lipoprotein-cholesterol levels but higher total cholesterol and cholesterol ester content in liver when compared with wild type mice(P＜0.05).The expression of SR-B1 mRNA in liver of CETP transgenic mice was significantly lower as compared with the control group(P＜0.05).

Objective To investigate the levels of cancer related fatigue (CRF) and the correlations between CRF and serum inflammatory factors and hypothalamus-pituitary-adrenal (HPA) axis in patients with gastrointestinal cancer.Methods The CRF level was assessed by brief fatigue inventory (BFI).The level of C-reactive protein (CRP) was measured by immunoturbidimetry, and the level of cortisol was measured by electrochemiluminesence.The levels of interleukin (IL)-6, IL-1β, tumor necrosis factor-α (TNF-α), adrenocorticotropic hormone (ACTH) and norepinephrine (NE) were measured by enzyme-linked immunosorbent assay (ELISA).Results The average total score of CRF was 3.15±1.93, and the degree was mild to moderate, which was positively correlated with the CRP (r=0.321, P=0.000), TNF-α (r=0.265, P=0.000), NE (r=0.174, P=0.015) and ACTH (r=0.257, P=0.000), but was not correlated with the cortisol (r=0.033, P=0.652).Eastern Cooperative Oncology Group (ECOG) score (t=8.081, P=0.000), education (t=-4.244, P=0.000), treatment (t=4.563, P=0.000), time from diagnosis to sampling (t=3.453, P=0.001) and CRP (t=2.837, P=0.006) were important factors of CRF.Conclusion The CRF status is common in gastrointestinal cancer patients.The CRF is correlated with the NE and ACTH of HPA axis.Medical staff should pay attention to the inflammatory factors and hormone levels to improve the fatigue status and the quality of patients.

Objective To investigate the variability in biochemistry in cerebral palsy children who accepted rehabilitation exercise. Methods From March to November, 2015, 24 children with cerebral palsy were measured the levels of hemoglobin, creatine kinase and blood urea before treatment. They all accepted routine rehabilitation exercise, and were measured hemoglobin, creatine kinase and blood urea again one week after treatment as the load increased, and six weeks after treatment at the end of exercise. They were assessed with Gross Motor Function Measure (GMFM-88) before and six weeks after treatment. Results The scores of GMFM-88 improved after treat-ment (t>7.12, P<0.001). Hemoglobin decreased (P<0.01), while creatine kinase and blood urea increased one week after treatment (P<0.01). Hemoglobin and blood urea came back to the levels before treatment six weeks after treatment (P?0.05), while creatine kinase reduced (P<0.01), but was still more than that before treatment (P<0.01). Conclusion Hemoglobin, creatine kinase and blood urea may respond the train-ing situation of cycle and the stage of exercise, and indicate the effects of load in the cerebral palsy children accepted rehabilitation exercise.

Objective:To explore the effect of Angelica Sinensis Volatile Oil on the mi RNA expression profiling in myocardium tissue of the spontaneously hypertensive rats (SHRs),and to clarify the possible mechanism of Angelica Sinensis Volatile Oil.Methods:All the SHRs were divided into Angelica group,model group and captopril group,and other Wistar rats with the same age were selected as control group.The non-invasive systolic blood pressure was detected;after 4 weeks of administration,the changes of miRNA expression profiling in myocardium tissue were measured by Affymetrix miRNA 4.0 Array.KEGG analysis was used to identify the target genes. Results:Compared with control group,the systolic blood pressure of the rats in model group was significantly decreased (P < 0.05)before treatment.4 weeks after administration,compared with model group,the systolic blood pressure of the rats in Angelica group was significantly decreased (P <0.05).Compared with model group, 29 differential miRNAs of rats in Angelica group were found (P < 0.05 ), with 13 up-regulated miRNAs and 16 down-regulated miRNAs.The KEGG analysis results showed that miR-19a,let-7i,and miR-181c were related to insulin signaling pathways; let-7i, miR-181a, and miR-455 were related to VEGF signaling pathways;miR-122,miR-181a, miR-200b, miR-181c, let-7i, and miR-19a were related to apoptosis (P < 0.05 ). Conclusion:Angelica Sinensis Volatile Oil can decrease the blood pressure in SHRs and it can regulate the blood pressure by regulating the miRNA related to insulin signaling pathways and VEGF signaling pathways.

Objective To explore the high-risk factors,clinical characteristics,therapy and prognosis of invasive fungal infection (IFI)in patients underwent allogeneic haemopoietic stem cell transplantation (AlloHSCT). Methods One hundred patients underwent Allo-HSCT at our department from March 2002 to July 2010 were analyzed retrospectively,among whom 26 patients had invasive fungal infection(IFI). Seven patients had pulmonary IFI before allo-HSCT, 14 patients had pulmonary IFI after allo-HSCT,3 patients had respiratory tract system IFI, and 2 patients had intestinal IFI. We observed the occurrence of Graft-versus-host disease (GVHD) ,cytomegalovirus( CMV )infection, Lymphocyte subsets and chronic basic diseases in patients with IFI. The twenty six cases were divided into two groups: experience therapy group with 12 cases and preemption therapy group with 14 cases. Results Among 26 patients with IFI,20 cases suffered from GVHD,6 cases had CMV infection,19 cases had low cellular immune function simultaneously. 1 case had diabetes,3 patients had pulmonary tuberculosis and 1 case had bronchiectasis as complications. In experience therapy groupe: 8 cases (67%)recovered completely but 1 case(8% )suffered from progressive infection. In preemption therapy groupe:3 cases ( 21% ) recovered completely but 5 cases ( 36% ) suffered from progressive infection. Conclusion Clinician should pay close attention to the patients with high-risk factors of IFI after allo-HSCT.