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.

Objective To observe the effects of neuromuscular activation (Neurac) training on balance and walking in children with hemiplegic cerebral palsy. Methods 30 children with hemiplegic cerebral palsy from March to October, 2015 were divided into observation group (n=15) and control group (n=15). Both groups received routine rehabilitation, while the observation group received 30-minute Neurac training in addition, 5 times a week, for 3 months. They were assessed with D and E domains of Gross Motor Function Measure 88 (GM-FM-88), balance of Fugl-Meyer Assessment and footprints analysis before and after treatment. Results The scores of D and E domains of GMFM-88 and balance of Fugl-Meyer Assessment, and the step length, step width and velocity improved in both groups after training (t>7.31, P<0.001), especially in the observation group (t>2.08, P<0.05). Conclusion Neuroac training can further promote the recovery of gross motor function, balance and walking in children with hemiplegic cerebral palsy.

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.

Objective To investigate the analgesic effects of intraperitoneal lithium chloride injection on radicular pain behaviors in rats.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion(CCD) ,40 male SD rats were randomly divided into model group and Sham-operation group (group S, n= 12) of radicular pain were established.The rats in the model group were subdivided randomly into Control group(group C, n= 12) ,Early treatment group(group E, n=8) and Later treatment group(group L, n= 8 ).Rats in group E were intraperitoneal injected with lithium chloride once daily on day 2 ～ 4 after CCD respectively,while rats in L,group C and S treated with Vehicle(0.9% NaCl).Rats in L group were intraperitoneal treated with lithium chloride on day 12 ～ 14 after CCD respectively,while rats in E,group C and S received Vehicle.The pain ethology indexes such as paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before operation and day 1,4,7,10, 14, 17 and 21 after operation.Results Compared to S group and preoperative level, PWMT and PWTL decreased at Day 1 postoperative in group C (P＜0.05).At day 4 after the operation,compared with group C(7.712 ±0.237)g and (8.190 ±0.382) s,PWMT and PWTL of E group increased to ( 14.607 ± 0,280) g and ( 19.940 ± 0.933 ) s (P ＜ 0.05 ) after intraperitoneal injected lithium chloride.At day 14, compared with group C ( 6.788 ± 0.331 ) g and ( 7.301 ± 0.481 ) s, PWMT and PWTL of group L increased to ( 11.700 ± 0.379) 8 and ( 18.524 ± 1.060) s (P ＜ 0.05 ).This analgesic effect of lithium chloride continued to exist at day 21.However, there was still a significant difference between S group and E,group L(P＜0.05).Conclusion Intraperitoneal lithium chloride injection alleviates pain behavior on radicular pain in rats.

Objective To investigate the effects of intrathecally coadministered dexamethasone and spironolactone in trathecally on radicular pain behaviors.Methods Using rat model of radicular pain induced by chronic compression of dorsal root ganglion (CCD) ,48 male SD rats successfully received intrathecal catheter implantation and without motor dysfunction were randomly divided into Sham-operation group (Sham group, n= 12),Control group ( C group, n = 12 ), Dexamethasone group ( D group, n = 8 ), Spironolactone group ( S group, n = 8 )and Dexamethasone plus spironolactone group (DS group, n=8).Rats in D group,S group or DS group were intrathecally treated with dexamethasone 4 μg, spironolactone 3 μg or dexamethasone 4 μg plus spironolactone 3 μg twice daily on day 2 ～4 after CCD respectively,while rats in C and Sham group received 10μl 10% alcohol.Paw withdrawal mechanical threshold(PWMT) and paw withdrawal thermal latency (PWTL) were tested on day 1 before CCD and day 1,4,7,10,14,17 and 21 after CCD.Results Compared with Sham group, both PWMT and PWTL were significantly decreased after CCD surgery on the ipsilateral side(P＜0.01 ＝.Intrathecally administrated with dexamethasone significantly improved pain behaviors (P＜0.01 ＝ and these therapeutic effects lasted up to 10 days after CCD surgery.As with dexamethasone,intrathecal spironolactone also significantly attenuated PWMT (P＜0.01 ＝ and PWTL (P＜0.01 ＝ and the change lasted up to 7 days after CCD surgery.Coadministration spironolactone and dexamethasone exhibited significant synergies( PWMT: ( 13.52 ± 0.72) g vs ( 11.58 ± 1.38 ) g, P ＜0.01; PWTL: ( 19.63 ± 1.68) s vs ( 14.14 ± 1.52) s, P ＜ 0.01 ＝.These effects lasted up to at least 10 days.Conclusion Both dexamethasone and spironolactone intrathecally have therapeutic effects on radicular pain behaviors, combination injection of these two drugs could generate significant synergies.

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.

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.